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Announces the appointment of Gunnar Kvalheim and J. Graham Sharp as co-editors of "Cytotherapy" journal. Career highlights in the field of stem cell transplantation; Educational background; Responsibilities the position require.

GvHD, the most important cause of morbidity and mortality after allogeneic stem cell transplantation, depends primarily on the ability of a donor T-cell subset to react to immunogenic host Ag. Recently developed culture conditions and treatment strategies may bring us closer to the selective elimination of such alloreactive T cells, often considered the holy grail of transplantation. Among the various therapeutic modalities, photodynamic therapy (PDT) offers a biological and global approach to the eradication of unwanted allo-activated T cells by combining mitochondrial targeting, P-glycoprotein inhibition and reactive oxygen species production. Indeed, the high potency of PDT against malignant cells has been harnessed to exert selective and extensive elimination of alloreactive T-cell subsets mediating GvHD, while preserving resting T cells with the ability to reconstitute the immune system for GvL activity and prevent or suppress viruses and fungi. The present paper reviews the basis of the PDT strategy, and the methodology employed. In vitro and in vivo studies that formed the proof of principle as a basis for human studies to investigate the clinical potential of PDT in the context of GvHD will be presented together with insights into future clinical applications of this versatile treatment platform. [ABSTRACT FROM AUTHOR]

Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice for many hematologic malignancies and inherited disorders of the hematopoietic system. Ex vivo T-cell depletion (TCD) of the graft and post-transplantation immunosuppression efficiently prevents the development of GvHD in no- MHC-identical settings. However, the consequence of these non-specific strategies is a long-lasting immunodeficiency associated with increased incidence of disease relapse, graft rejection and reactivation of viral infections. Donor lymphocyte infusion, which is used for treating leukemic relapse after allogeneic HSCT, can result in severe GvHD. Several strategies are being optimized specifically to inactivate anti-host T cells while preserving anti-leukemic or anti-microbial immunocompetence. Based on the ex vivo or in vivo elimination of anti-host T cells, or on the modulation of their anti-host activity, these approaches, which have been explored extensively in pre-clinical studies and tested in some preliminary clinical trials, are discussed in this paper. [ABSTRACT FROM AUTHOR]

Focuses on issues concerning cell therapy. Requirements for stem cells in clinical trials; Potential of cell-mediated therapy to direct potent and specific immune attack on malignant cells; Importance of first-class translational research in filling the gap between basic science and clinical treatment.

The concept of particulates, while common to many in the pharmaceutical and blood transfusion disciplines, represents a distinct challenge in the field of cellular therapy. With newly discovered products advancing through clinical trials, the focus has shifted to ensuring products are manufactured in a reliable and safe manner. Given the unique manufacturing processes and resulting products (i.e. the cell being the active ingredient of the product), the way in which particulates are viewed and subsequently tested needs to be reviewed. No specific test or method for particulates will apply to all products, and guidance documents will be generated over time as more cell therapy products are approved. The details of the processes, testing methods used and acceptance criteria established for particulates will play a major role in generating the guidance documents. This will ultimately allow for the manufacture and administration of safe and effective products without thwarting advancement of the cellular therapy field. The intent of this review is to bring awareness to the topic of particulates with respect to cell therapy, and encourage a more open dialog and exchange of examples within the industry. We have reviewed the concept of particulates, where they originate and how they are introduced to cell therapy products, and the current methods available for their detection. We have also reviewed the relevance of current guidance documents and present potential strategies to move forward and address and control unwanted contaminating particulates in cell therapy products. [ABSTRACT FROM AUTHOR]

Background aims. Immunomagnetic cell selection (ICS) of CD34+ cells is being used increasingly in allogeneic transplantation in order to reduce T-cell quantity. The aim of this study was to evaluate an automated washing protocol before immunomagnetic selection. Methods. The automated method was compared with a conventional washing procedure. In the study group the cell processing using the automated procedure, both before and after antibody incubation, was performed with a Sepax S-100 device. The efficacy of the automated procedure was compared with the control group, where washing were performed using a standard method. Results. The results obtained after pre-incubation washing performed using the automated system showed a total nucleated cell (NC) and CD34+ cell recovery of 84.87% (71.80-105, SD 8.62; range, standard deviation) and 83.45% (47-109, SD 16.12), respectively. The NC and CD34+ cell recovery after the pre-incubation washing cycle was performed using the standard method was 75.54% (38.36-97.76, SD 22.5) and 61.51% (30.87-81.79, SD 19.3), respectively. The CD34+ cell recovery after ICS was 51.27% (13.77-98.82, SD 24.97) and 48.89% (15.57-88.24, SD 25.91) for group 1 and group 2, respectively. The average purity in group 1 was 86.46% (67.4-96.10, SD 13.07) and in group 2 84.97% (58.1-97.8, SD 15.58). Conclusions. The efficacy of the ICS led to an optimal purity without affecting cell recovery, which was higher in group 1. Overall, our data suggest that the automated method is suitable for washing hematopoietic progenitor cell apheresis (HPC-A) concentrates before immunomagnetic cell selection in daily clinical routines. [ABSTRACT FROM AUTHOR]

The prognosis of patients with acute myeloid leukemia (AML) remains dismal, with a 5-year overall survival rate of only 5.2% for the continuously growing subgroup of AML patients older than 65 years. These patients are generally not considered eligible for intensive chemotherapy and/or allogeneic hematopoietic stem cell transplantation because of high treatment-related morbidity and mortality, emphasizing the need for novel, less toxic, treatment alternatives. It is within this context that immunotherapy has gained attention in recent years. In this review, we focus on the use of dendritic cell (DC) vaccines for immunotherapy of AML. DC are central orchestrators of the immune system, bridging innate and adaptive immunity and critical to the induction of anti-leukemic immunity. We discuss the rationale and basic principles of DC-based therapy for AML and review the clinical experience that has been obtained so far with this form of immunotherapy for patients with AML. [ABSTRACT FROM AUTHOR]

The article focuses on the preparation of autologous bone marrow (BM) mononucleated cells (MNC) for the clinical treatment of peripheral arterial disease and acute myocardial infarction (AMI). Data from a meta-analysis and systematic review of the literature suggest the safety of bone marrow cell (BMC) transplantation and the association with modest improvements in remodeling in AMI patients. It adds that the quality of the starting material is the crucial aspect in the clinical treatment.

Background aims. Recalcitrant diabetic wounds are not responsive to the most common treatments. Bone marrow-derived stem cell transplantation is used for the healing of chronic lower extremity wounds. Methods. We report on the treatment of eight patients with aggressive, refractory diabetic wounds. The marrow-derived cells were injected/applied topically into the wound along with platelets, fibrin glue and bone marrow-impregnated collagen matrix. Results. Four weeks after treatment, the wound was completely closed in three patients and significantly reduced in the remaining five patients. Conclusions. Our study suggests that the combination of the components mentioned can be used safely in order to synergize the effect of chronic wound healing. [ABSTRACT FROM AUTHOR]

Transplantation is the treatment of choice for many diseases that result in organ failure, but its success is limited by organ rejection. Stem cell therapy has emerged in the last years as a promising strategy for the induction of tolerance after organ transplantation. Here we discuss the ability of different stem cell types, in particular mesenchymal stromal cells, neuronal stem/progenitor cells, hematopoietic stem cells and embryonic stem cells, to modulate the immune response and induce peripheral or central tolerance. These stem cells have been studied to explore tolerance induction to several transplanted organs, such as heart, liver and kidney. Different strategies, including approaches to generating tolerance in islet transplantation, are discussed here. [ABSTRACT FROM AUTHOR]

Background aims. Transplantation of mesenchymal stromal cells (MSC) derived from bone marrow (BM) or adipose tissue is expected to become a cell therapy for stroke. The present study compared the therapeutic potential of adipose-derived stem cells (ASC) with that of BM-derived stem cells (BMSC) in a murine stroke model. Methods. ASC and BMSC were isolated from age-matched C57BL/6J mice. These MSC were analyzed for growth kinetics and their capacity to secrete trophic factors and differentiate toward neural and vascular cell lineages in vitro. For in vivo study, ASC or BMSC were administrated intravenously into recipient mice (1 ×× 105 cells/mouse) soon after reperfusion following a 90-min middle cerebral artery occlusion. Neurologic deficits, the degree of infarction, expression of factors in the brain, and the fate of the injected cells were observed. Results. ASC showed higher proliferative activity with greater production of vascular endothelial cell growth factor (VEGF) and hepatocyte growth factor (HGF) than BMSC. Furthermore, in vitro conditions allowed ASC to differentiate into neural, glial and vascular endothelial cells. ASC administration showed remarkable attenuation of ischemic damage, although the ASC were not yet fully incorporated into the infarct area. Nonetheless, the expression of HGF and angiopoietin-1 in ischemic brain tissue was significantly increased in ASC-treated mice compared with the BMSC group. Conclusions. Compared with BMSC, ASC have great advantages for cell preparation because of easier and safer access to adipose tissue. Taken together, our findings suggest that ASC would be a more preferable source for cell therapy for brain ischemia than BMSC. [ABSTRACT FROM AUTHOR]

Background aims. Enumeration of CD34++ cells in leukocyte-rich cell suspensions is important for clinical decision-making in stem cell transplantation. Single-platform flow cytometry assays offer the significant advantages of speed and reproducibility, and have therefore become the gold standard in stem cell enumeration. The clinical community has recently defined the need for stem cell enumeration kits that incorporate viability dyes. The purpose of this study was to evaluate a novel assay, BD Biosciences'' (BD) stem cell enumeration kit (SCE kit‡‡), in relation to Beckman Coulter's (BC) commercially available BC Stem-Kit™™. Methods. Fresh/freeze-thawed samples from leukapheresis, bone marrow and cord blood, and fresh normal/mobilized blood, were analyzed with both assays (simultaneous detection of side/forward scatter and three fluorescence signals) on two flow cytometry platforms, BD FACSCanto II and BD FACSCalibur. Results. Results from both assays were highly congruent, with an overall r2 ≥≥ 0.99 (all specimen types included), a linear correlation across all CD34++ cell frequencies and concentrations, and an almost ideal steepness of the trend line. Conclusions. Both assays functioned reliably. Being based on single-platform International Society of Hematotherapy and Graft Engineering (ISHAGE) guidelines and similar staining methods, both assays essentially come to identical results. For most specimen types, the viability of CD34++ cells was equal to overall leukocyte viability. In summary, in the hands of an experienced technician, the BD™™ SCE kit and the BC Stem-Kit are equivalent. The infrequent user might derive benefit from the fact that counting spheres are pre-pipetted into the Trucount tube for the SCE kit, making this assay less susceptible to pipetting inaccuracy. [ABSTRACT FROM AUTHOR]

Background aims. Spinal cord injury is common among young subjects involved in motor vehicle accidents. Mechanisms and attempts to reverse post-traumatic pathophysiologic consequences are still being investigated. Unfortunately no effective and well-established treatment modality has been developed so far. The regeneration capability of the human nervous system following an injury is highly limited. Methods. The study involved four patients (two male, two female) who had suffered spinal cord injury as a result of various types of trauma. On neurologic examination, all the patients were determined to be in American Spinal Injury Association (ASIA) grade A. All patients were treated with decompression, stabilization and fusion for vertebral trauma anteriorly, as well as intralesional implantation of cellular bone marrow concentrates using a posterior approach 1 month after the first operation. The patients were then treated and followed-up in the physical rehabilitation clinic. Results. At the end of the post-operative 1-year follow-up, two of the patients were classified as ASIA C while one was classified as ASIA B. One patient showed no neurologic change; none of the patients suffered from any complications or adverse effects as a result of intralesional application of bone marrow cells. Conclusions. The results of this experimental study show the potential contribution of intralesional implantation of bone marrow to neuronal regeneration in the injured spinal cord, with neuronal changes. In light of the results of this experimental study, the potential for regenerative treatment in injuries of the human spinal cord is no longer a speculation but an observation. [ABSTRACT FROM AUTHOR]

Today, many patients suffer from acute liver failure and hepatoma. This is an area of high unmet clinical need as these conditions are associated with very high mortality. There is an urgent need to develop techniques that will enable liver tissue engineering or generate a bioartificial liver, which will maintain or improve liver function or offer the possibility of liver replacement. Liver tissue engineering is an innovative way of constructing an implantable liver and has the potential to alleviate the shortage of organ donors for orthotopic liver transplantation. In this review we describe, from an engineering perspective, progress in the field of liver tissue engineering, including three main aspects involving cell sources, scaffolds and vascularization. [ABSTRACT FROM AUTHOR]

Background aims. Spinal cord injury is a devastating injury that impacts drastically on the victim's quality of life. Stem cells have been proposed as a therapeutic strategy. Neural stem (NS) cells have been harvested from embryonic mouse forebrain and cultured as adherent cells. These NS cells express markers of neurogenic radial glia. Methods. Mouse NS cells expressing green fluorescent protein (GFP) were transplanted into immunosupressed rat spinal cords following moderate contusion injury at T9. Animals were left for 2 and 6 weeks then spinal cords were fixed, cryosectioned and analyzed. Stereologic methods were used to estimate the volume and cellular environment of the lesions. Engraftment, migration and differentiation of NS cells were also examined. Results. NS cells integrated well into host tissue and appeared to migrate toward the lesion site. They expressed markers of neurons, astrocytes and oligodendrocytes at 2 weeks post-transplantation and markers of neurons and astrocytes at the 6-week time-point. NS cells appeared to have a similar morphologic phenotype to radial glia, in particular at the pial surface. Conclusions. Although no functional recovery was observed using the Basso Beattie Bresnahan (BBB) locomotor rating scale, NS cells are a potential cellular therapy for treatment of injured spinal cord. They may be used as delivery vehicles for therapeutic proteins because they show an ability to migrate toward the site of a lesion. They may also be used to replace lost or damaged neurons and oligodendrocytes. [ABSTRACT FROM AUTHOR]

Immunotherapy has become an important part of hematopoietic stem cell (HSC) transplantation and cancer therapy. Regenerative and reparative properties of somatic cell-based therapies hold tremendous promise for repairing injured tissue, preventing and reversing damage to organs, and restoring balance to compromised immune systems. The principles and practices of the diverse aspects of immune therapy for cancer, HSC transplantation and regenerative medicine have many commonalities. This meeting report summarizes a workshop sponsored by the National Heart, Lung and Blood Institute (NHLBI) and Production Assistance for Cellular Therapies (PACT), held on 23–24 April 2009 at the National Institutes of Health (NIH, USA). A series of scientific sessions and speakers highlighted key aspects of the latest scientific, clinical and technologic developments in cell therapy, involving a unique set of cell products with a special emphasis on converging concepts in these fields. [ABSTRACT FROM AUTHOR]

As a promising concept for regeneration medicine, endothelial progenitor cell (EPC) therapy represents a novel strategy for a variety of diseases. Increasing evidence suggests that the spleen, a traditionally dispensable organ, acts as a major reservoir during EPC trafficking and plays an important role regarding the modulation of circulating EPC kinetics. Moreover, infusion of splenic EPC can restore endothelial function and promote neovascularization, indicating an available resource for EPC transplantation. Thus a discussion of the role of the spleen with respect to EPC may provide novel information for management of EPC therapy. [ABSTRACT FROM AUTHOR]

Background aims Mesenchymal stromal cells (MSC) have been used in a wide variety of pre-clinical experiments and in an increasing number of human clinical trials. Although many of these studies have shown different levels of engraftment, the exact fate of MSC after transplantation and the tissue response to their engraftment have not been investigated in detail. In the present work we studied the distribution of human MSC in a rat hind limb ischemic injury model immediately after transplantation and also analyzed the recipient tissue response to transplanted cells. Methods We tracked the in vivo fate of the transplanted MSC utilizing bioluminescence imaging, fluorescence microscopy and gene/protein expression analysis in a rat hind limb ischemia model. We also monitored the viability of transplanted cells by graft versus recipient expression analysis and determined the angiogenic and proliferative effect of transplantation by histologic staining. Results According to imaging analysis only a small portion of cells persisted for an extended period of time at the site of injury. Interestingly, recipient versus graft expression studies showed increased synthesis of rat-origin angiogenic factors and no human-origin mRNA or protein synthesis in transplanted tissues. More importantly, despite the lack of robust engraftment or growth factor secretion the transplantation procedure exerted a significant pro-angiogenic and pro-proliferative effect, which was mediated by angiogenic and mitogenic signaling pathways. Conclusions Our results show an immediate temporal tissue effect in response to MSC transplantation that may represent a novel indirect paracrine mechanism for the beneficial effects of cell transplantation observed in injured tissues. [ABSTRACT FROM AUTHOR]

Background Successful clinical development of novel cellular therapeutics requires the evaluation of clinical acute toxicity endpoints in scoring patient adverse events (AE) contributing to dose-limiting toxicity (DLT) for establishment of the maximum-tolerated dose (MTD). However, many clinical pathology parameters are not routinely evaluated in pre-clinical safety testing. The objective of this pre-clinical study was to investigate thoroughly the acute toxicity of single- and multiple-dose administrations of allogeneic multipotent adult progenitor cells (MultiStem®), which represent a class of stromal stem cells with therapeutic potential. MethodsMultiStem were tested as an adjunct treatment in a rat myeloablative hematopoietic stem cell transplantation (HSCT) model for impact on clinical parameters, clinical chemistry, hematology, immunology and histopathology parameters. Animals received MultiStem in a single dose of 12.5 million cells/kg on day 2 after HSCT or in five infusions at this dose on days 2, 9, 16, 23 and 30. Controls received phosphate-buffered saline injections and all animals were killed on day 37. Results There were no significant differences between tests and controls regarding evaluation of respiratory distress upon infusion, clinical assessment and hematology and clinical chemistry analysis. Gross necropsy and histopathology analysis showed no organ profile alterations. There was no significant evidence for allogeneic antibody production or T-cell sensitization upon MultiStem infusion. Discussion These studies demonstrate the safety of administration of allogeneic stromal stem cells in repeat dosing regimens in bone marrow transplant settings, and define pre-clinical safety testing standards relevant to the development of cellular therapeutics using allogeneic adherent adult stem cells. [ABSTRACT FROM AUTHOR]

Background To study cell replacement therapy using embryonic stem (ES) cells in mice, avoiding immune rejection and tracing the fate of transplanted cells are important issues. This study was carried out to isolate ES cells ubiquitously expressing enhanced green fluorescent protein (EGFP) and test the survival of these cells in allografts in the cochlea of inbred C57BL/6 mice. Methods Putative ES cells were isolated from blastocysts collected from C57BL/6-green mice ubiquitously expressing EGFP. Pluripotency of these cells was tested by expression of stem cell markers and in vitro differentiation of the cells into embryoid bodies. Isolated EGFP-transgenic ES cells were injected into the cochlea of deafened inbred C57BL/6 mice, and survival of transplanted cells was identified in histologic sections of the cochlea. Results Putative ES cells expressed cellular markers for ES cells, including alkaline phosphatase, Oct-4, Nanog and stage-specific embryonic antigen-1. These cells formed embryoid bodies in suspension cultures. Incorporation of transplanted cells was found at the area of spiral ganglion neurons, auditory nerve fibers reaching the organ of Corti and stria vascularis in the scala media. Grafted cells were also found at the location of inner hair cells underneath the tectorial membrane. Discussion The isolation of ES cells from the EGFP-transgenic mouse and transplantation into allogeneic inbred mice may be a useful means of studying cell therapy with respect to the ubiquitous and stable expression of EGFP and elimination of graft rejection. [ABSTRACT FROM AUTHOR]

Background Both bone marrow stromal cells (BMSC) and olfactory ensheathing cells (OEC) have been demonstrated experimentally as promising for therapy of spinal cord injury (SCI). However, clinical use may be constrained by the margin neuronal differentiation capacity of BMSC as well as the limited number of isolatable OEC. This study therefore tested the efficacy of co-grafting human BMSC and OEC in treating thoracic SCI. Methods Rat SCI models were created with cushion forces. OEC were labeled with Hoechst 33342 and BMSC with BrdU or GFP. BMSC, OEC and BMSC plus OEC were injected into the injured sites of rat spinal cords. Histologic, electrophysiologic and functional approaches were applied to assess the effects of transplantation of these cell types. Results Behavioral evaluation showed an improvement in animals with all cell-based treatments. The co-graft led to significantly higher gait scaling. The latency of transcranial magnetic motor-evoked potential (tcMMEP) responses was also better restored in the co-graft group. Larger numbers and sizes of axon bundles through the transitional zone between the normal and injured regions were observed in the co-graft animals in comparison with all other animals. Transplanted bone marrow stromal cells were identified as neurofilament-positive in the co-grafted animals although the number of glial fibrillary acidic protein-positive cells remained the same in all groups. Discussion Taken together, our results suggest that the combined use of BMSC and OEC may provide an improved approach for the treatment of SCI. [ABSTRACT FROM AUTHOR]

Background Mesenchymal stem cells (MSC) have recently been shown to possess immunomodulatory properties in vitro and in vivo. The present study aimed to investigate the regulatory effect of MSC transplantation on the immuno-inflammatory response in myocardial infarction (MI). Methods MI was induced in Sprague-Dawley rats by left anterior descending coronary artery ligation, and the animals were randomly assigned into the following three groups: sham ( n=8); phosphate-buffered saline (PBS) injected (MI+PBS, n=8); and MSC transplantation (MI+MSC, n=8). BrdU-labeled MSC or PBS was transplanted into peri-infarct myocardium by direct myocardial injection. At 1 and 28 days post-transplantation, cardiac function was evaluated by echocardiography. Transplanted cells were investigated through immunohistochemistry. Lymphocyte cytotoxic activity was evaluated with the crystal violet method. The activity of NF-κB and protein expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-10 in myocardium were assessed by immunohistochemistry and Western blot. Results Echocardiographic examination revealed that the MSC transplantation prevented left ventricular dilation and dysfunction at 28 days after the operation. BrdU-stained cells were found living in host heart 4 weeks after transplantation. MSC transplantation attenuated the cytotoxic activity of spleen lymphocytes. Transplantation of MSC inhibited the activity of NF-κB, attenuated the protein production of TNF-α and IL-6, and increased the expression of IL-10 in peri-infarct myocardium. Discussion MSC transplantation modulated the immuno-inflammatory response in MI. The immuno-inflammatory regulatory effect of MSC transplantation might partly account for the cardiac protection in myocardial infarction. [ABSTRACT FROM AUTHOR]

Background Autologous PBSC transplantation is the standard care for patients with multiple myeloma. The most common regimen used to mobilize PBSC consists of CY and G-CSF. Methods We retrospectively analyzed the efficacy and toxicity of two regimens of CY for PBSC mobilization: low-dose CY (1-2 g/m2, LD-CY, n=61) plus G-CSF, and intermediate-dose CY (3-4 g/m2, ID-CY, n=26) plus G-CSF. Results In the LD-CY group, 5.17 (0.23-17.3)×106 CD34+ cells/kg, and in the ID-CY group 7.71 (0.08-26.4)×106 CD34+ cells/kg (P=0.018), were collected. Although ≥2×106/kg CD34+ cells were collected in 89% of the LD-CY group and 92% of the ID-CY group, this was achieved after a single leukapheresis in 54% of the LD-CY group and 92% of the ID-CY group (P=0.0001). Patients who are to have tandem autologous PBSC transplants require ≥4×106/kg CD34+ cells. This was achieved in only 65% patients in the LD-CY group but 88% in the ID-CY group (P=0.05). Among patients who had not had prior melphalan and/or >12 months of prior treatment, 74% in the LD-CY group and 100% in ID-CY group mobilized ≥4x106/kg CD34+ cells. Febrile neutropenia was more frequent in the ID-CY group (38% vs. 13%). Discussion In conclusion, compared with LD-CY, patients receiving ID-CY were more likely to collect a total CD34+ cell number adequate for tandem autologous PBSC transplantation. The increased toxicity was manageable and considered acceptable. [ABSTRACT FROM AUTHOR]

Not all patients with Hodgkin's disease (HD) respond to standard chemotherapy and/or radiation, and hematopoietic stem-cell transplantation has been gaining increasing acceptance in the management of HD. Phase II and, to a lesser extent, phase III studies of high-dose chemotherapy and autologous stem cell transplantation carried out at multiple institutions worldwide have proven the feasibility of the procedure and provided extended progression-free survival (and possibly cure) in a sizable number of patients with relapsed or refractory HD. Prognostic factors have been identified by multiple investigators (with response to chemotherapy being the most impressive one) and may ultimately allow a risk-adapted strategy. While early and late treatment-related morbidity and mortality remains an issue, with current supportive care modalities most patients tolerate this procedure with only minor or manageable complications. Disease recurrence remains a problem in many patients, and this can unfortunately occur as late as six or seven years after a seemingly successful transplant. New chemotherapeutic agents and strategies (such as post-transplant maintenance and possibly immunomodulation) will be required to successfully tackle this issue. Allogeneic stem-cell transplantation from HLA-compatible donors has yielded largely unsatisfactory results in the published studies in the literature, despite favorable results in a small minority of patients. Recently, however, newer approaches and strategies (such as the introduction of reduced-intensity, purine analog-based conditioning regimens and possibly cellular immunotherapy in the form of donor lymphocyte infusions) have provided very encouraging early results and seem to brighten the outlook for this procedure. [ABSTRACT FROM AUTHOR]

Background: We report our updated experience of allogeneic transplantation in lympho-proliferative disorders using a reduced-intensity conditioning regimen combining BEAM (plus fludarabine in three cases) with pre-transplant CAMPATH. Post-transplant donor lymphocytes have been infused for persisting disease or relapse, and both chimerism and minimal residual disease have been monitored utilizing molecular techniques. Methods: Thirty patients with median age 47.6 years underwent allogeneic transplantation for relapsed or high-risk lymphoproliferative disease using HLA-identical (sibling n = 25, unrelated n = 2) or one antigen mismatched sibling donors (n = 3). Twenty-one had NHL, three had HD and six had CLL/PLL. Stem-cell source was PBSC (n = 24), BM (n = 5) or both (n = 1) with a median CD34 dose of 4.5 × 10[sup 6]/kg. GvHD prophylaxis was with CYA and MTX. Results: Engraftment was prompt in the majority of patients, with a median of 15 days to both ANC > 0.5 and platelets > 20. There have been three transplant-related deaths secondary to viral pneumonitis or bacterial pneumonia. Seven patients developed Grade I–II acute GvHD post-transplant. Of 28 evaluable patients, 18 achieved a CR at assessment 2–3 months post-transplant and a further patient converted from PR to CR following DLI, to give an overall CR rate of 68%. Three patients had early progressive disease and six have relapsed from CR or progressed from PR (two of whom have achieved CR following DLI therapy). Overall survival is 67% and event-free survival 48% at 3 years. With a median follow-up of 1.3 years 57% of patients are currently alive and lymphoma-free. A molecular remission has been achieved in nine of 12 informative patients. Discussion: These encouraging results show that this reduced-intensity conditioning regimen is effective, with a low-toxicity profile compared with conventional TBI-based conditioning, and certainly merits further evaluation in this setting. [ABSTRACT FROM AUTHOR]

Background: We have investigated a novel nonmyeloablative conditioning regimen in 44 patients with hematological malignancies. The median patient age was 41 years. Many of the patients had high-risk features, including 19 patients with a previous failed transplant. Methods: Recipient conditioning consisted of CAMPATH-1H 20 mg/day on Days -8 to -4, fludarabine 30 mg/m[sup 2] on Days -7 to -3 and melphalan 140 mg/m[sup 2] on Day -2. Thirty-six recipients received unmanipulated G-CSF mobilized PBSC from HLA identical siblings and eight received unmanipulated BM from MUD. GvHD prophylaxis was with CYA alone for 38 patients and CYA plus MTX for six sibling recipients. Results: Forty-two of the 43 evaluable patients had sustained engraftment. Results of chimerism analysis using microsatellite PCR indicate that 18 of 31 patients studied were full donor chimeras, while the other patients were mixed chimeras in one or more lineages. At a median follow-up of 9 months (range, 3–29 months) 33 patients remain alive in CR, or with no evidence of disease progression. Seven patients relapsed or progressed post-transplant and four of them subsequently died. Four patients died from regimen-related complications. There were no cases of Grades III–IV acute GvHD. Only two patients developed Grade II acute GvHD and only one had chronic GvHD. The estimated probability of non-relapse mortality at 1 year was 11%. Results: Although longer follow-up is needed to establish the long-term remission rates, this study demonstrates that this nonmyeloablative preparative regimen is associated with durable engraftment, minimal toxicity and low incidence of GvHD. [ABSTRACT FROM AUTHOR]

Background We report a routine flow cytometric (FACS) approach to quantify circulating leukocytes (NC) in myeloablated patients before and during regeneration after allogeneic transplantation of either whole bone marrow (BM) or of highly purified (> 99%) blood-derived CD34 + cells (PBSC). Methods Blood samples were analyzed daily between infusion of the transplant and hematopoietic reconstitution. Significant differences in the composition of NC types and CD34 + cells were observed between the two CD34 sources. The detection threshold for NC was roughly 1 cell per μL blood. Results The cell nadir of < 100 NC/μL was reached on Day +4 (BM) and on day 0 (PBSC), when unusual CD34 + cells of recipient genotype were detected in all patients. They were not clonogenic, showed high CD34 expression, but were negative for CD45, CD38, CD33, CD50, HLA-DR and Stro-1. Between Days +5 and +16, the onset of hematopoietic reconstitution was clearly detectable in multi-parameter evaluation of the FACS data. This was a median of 3.5 days before NC increased above 200/μL blood and 4-10 days before granulocyte counts were > 500/μL. It was marked by the appearance of monocytes, immature (CD38 + ) granulocytes, and clonogenic donor CD34 + cells exhibited normal size and phenotype. Discussion We conclude that dynamic FACS analyses can reliably detect hematopoietic reconstitution, but also graft rejection, before a visible increase NC numbers. This may have considerable impact on clinical management strategies. [ABSTRACT FROM AUTHOR]

Background aims. We carried out a retrospective analysis of viability by diagnosis and dimethyl sulfoxide (DMSO) concentration in patients who had undergone autologous transplants using hematopoietic progenitor cells (HPC) after long-term storage (up to 17.8 years). Methods. Viability was tested using flow cytometry for HPC that were harvested and preserved using a controlled rate freezer and 5% or 10% DMSO with human serum albumin, then stored in liquid nitrogen. Data from 262 samples were analyzed (249 myeloma patients and 13 other diagnoses): 100 consecutively thawed samples with a storage time of <1 year (all 10% DMSO), 50 consecutive samples stored for 1-4.9 years (10% DMSO), 50 samples stored for 5-9 years (5% DMSO) and all samples stored and used for transplant after >9 years (60 samples, 5% DMSO; two samples, 10% DMSO). Results. No statistically significant difference in viability between the 5% DMSO and 10% DMSO groups was observed ( P = 0.08), so the 1-4.9 years and 5-9 years were combined and the three groups (<1 year, 1-9 years and >9 years) were compared using an anova test. There was no difference in viability based on cryostorage period ( P = 0.23) or between myeloma and other diagnoses ( P = 0.45). No difference was seen in time to White blood cell (WBC) engraftment ( P = 0.10) or to platelet engraftment between groups ( P = 0.52). Conclusions. These data suggest that long-term storage in 5% DMSO and human serum albumin is safe. [ABSTRACT FROM AUTHOR]

The article discusses the allogeneic bone marrow mesenchymal stromal cell-derived hepatocyte-like cell transplantation in homozygous familial hypercholesterolemia (FH). It defines FH as a monogenic autosomal disorder caused by loss of gene allele function encoding low-density lipoprotein (LDL) receptor. In the study conducted, human mesenchymal stromal cells (MSC) is found to have the ability to differentiate into hepatocyte-like cells that are functional.

The article discusses the standardization of cell analysis methods in clinical cellular therapy programs. Accordingly, the cell therapy programs were being developed that will also require standardization of the analytical methods used for quality assessment of the cell products. The external assurance programs for lymphocyte subset enumeration and cell enumeration have led to significant reductions in between laboratory variation.

Focuses on cellular therapy for diabetes mellitus. Obstacles to the attempts to broaden the applicability of promising results of transplantation of patients with type 1 diabetes mellitus; Prospect of identifying and culturing human stem or progenitor cells capable of ex vivo generation of islets; Possible sources of pancreatic precursor cells.