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Background. Mesenchymal stem cells (MSCs) have a therapeutic potential for the treatment of osteoarthritic (OA) joint pathology and pain. The aims of this study were to determine the influence of a passage number on the effects of MSCs on pain behaviour and cartilage and bone features in a rodent model of OA. Methods. Rats underwent either medial meniscal transection (MNX) or sham surgery under anaesthesia. Rats received intra-articular injection of either 1.5 × 106 late passage MSCs labelled with 10 μg/ml SiMAG, 1.5 × 106 late passage mesenchymal stem cells, the steroid Kenalog (200 μg/20 μL), 1.5 × 106 early passage MSCs, or serum-free media (SFM). Sham-operated rats received intra-articular injection of SFM. Pain behaviour was quantified until day 42 postmodel induction. Magnetic resonance imaging (MRI) was used to localise the labelled cells within the knee joint. Results. Late passage MSCs and Kenalog attenuated established pain behaviour in MNX rats, but did not alter MNX-induced joint pathology at the end of the study period. Early passage MSCs exacerbated MNX-induced pain behaviour for up to one week postinjection and did not alter joint pathology. Conclusion. Our data demonstrate for the first time the role of a passage number in influencing the therapeutic effects of MSCs in a model of OA pain.

This article analyses the sociopolitical interactions that shaped an early colonial union statute that constituted a milestone because of its early passage, draconian nature and wider influence in the British Empire. We analyse the interactions between the Governor on the one hand and local and international actors on the other, to create the first trade union law in Mauritius. Colonial Office officials pressured the Governor to overcome local resistance to legislation, but this opposition and his attitudes shaped the law's content.

The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, our understanding of the biology of the canine DP is extremely limited. The aim of this study was to elucidate molecular biological and immunohistochemical characteristics of canine DP cells and determine appropriate conditions for in vitro expansion. Histological investigation revealed that the canine DP expressed biomarkers of human and rodent DP, including alkaline phosphatase (ALP) and versican. When microdissected, canine DP, but not fibroblasts, strongly expressed the DP-related genes for alkaline phosphatase, Wnt inhibitory factor 1 and lymphoid enhancer-binding factor 1, confirming successful isolation. The growth rate of isolated canine DP cells was moderate in conventional culture conditions for rodent and human DP; however, AmnioMAX-C100 complete medium allowed more efficient cultivation. Dermal papilla marker gene expression was maintained in early passage cultured DP cells, but gradually lost after the third passage. Approaches to mimic the in vivo DP environment in culture, such as supplementation of keratinocyte-conditioned medium or use of extracellular matrix-coated dishes, moderately ameliorated loss of DP gene expression in canine DP cells. It is possible that constituent factors in AmnioMAX may influence culture. These findings suggested that further refinements of culture conditions may enable DP cell expansion without impairing intrinsic properties and, importantly, demonstrated that AmnioMAX-cultured early passage canine DP cells partly maintained the biological characteristics of in vivo canine DP cells. This study provides crucial information necessary for further optimization of culture conditions of canine DP. Resume La papille dermique (DP) joue un role essentiel dans la morphogenese et le cycle du follicule pileux. Toutefois, notre comprehension de la biologie de la DP canine est extremement limitee. L’objectif de cette etude etait de determiner les caracteristiques des cellules de DP canine ainsi que les conditions appropriees pour leur expansion in vitro. L’analyse histologique a revele que la DP canine exprimait les biomarqueurs de DP humaine et de rongeurs, regroupant la phosphatase alcaline (ALP) et Versican. Apres dissection, la DP canine mais pas les fibroblastes, exprimait fortement les genes lies a l’ALP de DP, WIF1(Wnt inhibitory factor 1) et LEF1 (Lymphoid enhancer-binding factor 1), confirmant le succes de l’isolement. Le taux de croissance des cellules de DP canine isolees etait modere sous des conditions de culture conventionnelle pour DP d’humains et de rongeurs. Cependant le milieu complet AmnioMAX-C100 a permis une culture plus efficace. L’expression du gene marqueur etait maintenue dans les premiers passages de cultures de cellules de DP mais etait progressivement perdue apres le troisieme passage. Les essais pour mimer l’environnement in vivo de DP en culture, comme la suplementation avec milieu de culture conditionnes pour keratinocytes ou l’utilisation de plaques revetues de matrices extracellulaires, ont permis une amelioration moderee de la perte de l’expression de gene de DP par les cellules canines de DP. Il est possible que les facteurs constituants de AmnioMAX influencent la culture. Ces resultats suggerent que des affinements supplementaires des conditions de culture pourraient permettre l’expansion des cellules de DP sans compromettre leurs proprietes intrinseques et surtout, que les premiers passages de culture de cellules de DP canines sur AmnioMAX conservaient partiellement les caracteristiques biologiques des cellules de DP canines in vivo. Cette etude fourni une information cruciale necessaire pour l’optimisation des conditions de futures cultures de DP canines. Resumen La papilla dermica (DP) juega un papel esencial en la morfogenesis y el ciclo del foliculo piloso. Sin embargo, nuestro entendimiento de la biologia del la DP canina es muy limitado. El proposito de este estudio fue elucidar las caracteristicas moleculares, biologicas e inmunohistoquimicas de las celulas de la DP y determinar la condiciones mas apropiadas para la expansion in vitro. La investigacion histologica revelo que la DP canina expresaba biomarcadores de la DP de humanos y de ratones, incluida la fosfatasa alcalina (ALP) y el versican. Cuando se utilizo microdiseccion, las celulas de la DP canina, pero no los fibroblastos, expresaron intensamente genes relacionadas con la DP como ALP, factor de inhibicion Wnt-1 (WIF-1) y el factor de estimulacion de union linfoide-1 (LEF-1), confirmando un aislamiento exitoso. El ritmo de crecimiento de las celulas aisladas de la DP canina fue moderado bajo condiciones convencionales de cultivo para celulas de la DP de ratones y de humanos, sin embargo, medio completo AmnioMAX-C100 permitio un cultivo mas eficiente. Se mantuvo la expresion de los marcadores de la DP en las celulas de los pases tempranos en cultivo, pero gradualmente se perdieron tras el tercer pase. Intentos de reproducir la condiciones del microambiente de la DP in vivo en cultivo, tal como el suplemento con medio condicionado para queratinocitos o el uso de placas cubiertas con matriz extracellular, ligeramente mejoraron la perdida de los genes de la DP en la celulas. Es posible que factores constitutivos del medio AmnioMAX influyan el cultivo. Estos hallazgos sugieren que es necesario mejorar las condiciones de cultivo para permitir la expansion de las celulas de la DP sin alterar las propiedades intrinsecas, y, lo que es mas importante, demostro que pases iniciales de celulas DP en cultivo con medo AmnioMAX mantenian parcialmente las caracteristicas de las celulas de la DP in vivo. Este estudio aporta informacion crucial necesaria para posterior optimizacion del las condiciones de cultivo de celulas de la DP canina. Zusammenfassung Die dermale Papille (DP) spielt eine entscheidende Rolle bei der Morphogenese des Haarfollikels und des Haarzyklus. Nichtsdestotrotz ist unsere Kenntnis der Biologie der caninen DP stark limitiert. Es war das Ziel dieser Studie, die molekularbiologischen und immunhistochemischen Merkmale der caninen DP Zellen zu beleuchten und geeignete Bedingungen fur ihre Entwicklung in vitro zu bestimmen. Die histologische Untersuchung zeigte die Expression von Biomarkern der Papilla des Menschen und der Nager, ebenso der alkalischen Phosphatase (ALP) und von Versican. Nach der Mikrodissektion bestand eine starke Expression der caninen DP (nicht aber der Fibroblasten) von DP-verwandten Genen ALP, Wnt Inhibitory Factor 1 (WIF1) und Lymphoid enhancer-binding factor 1 (LEF1), was eine erfolgreiche Isolierung bestatigte. Die Wachstumsrate von isolierten caninen DP-Zellen war bei konventionellen Kulturbedingungen fur die DP des Menschen und der Nager masig, AmnioMAX-C100 Komplettmedium erlaubte jedoch eine effizientere Kultivierung. Die Expression von DP Markergenen war in DP-Zellen der Fruhkultur erhalten, ging aber nach der dritten Passage graduell verloren. Methoden, die Umgebung von DP in vivo in der Kultur nachzuahmen, wie etwa die Supplementierung von Keratinozyten-konditioniertem Medium oder der Verwendung von extrazellularen mit Matrix ausgekleideten Gefasen, verringerte den Verlust der DP Genexpression in den caninen DP Zellen nur masig. Es besteht die Moglichkeit, dass Bestandteile von AmnioMAX die Kultur beeinflussen konnten. Diese Ergebnisse geben Hinweise darauf, dass eine weitere Vervollkommnung der Kulturbedingungen das Wachstum von DP Zellen ermoglichen konnte ohne ihre intrinsischen Eigenschaften zu beeintrachtigen und es wurde etwas sehr Wesentliches gezeigt, namlich dass caninen DP Zellen einer Fruhkultur in AmnioMAX teilweise die biologischen Charakteristika der caninen DP Zellen in vivo erhalten geblieben sind. Diese Studie liefert wichtige Informationen, die fur eine zukunftige Optimisierung der Kulturbedingungen fur canine DP Zellen essentiell sind.

Human embryonic stem cell (hESC) growth is dependent on various factors released by feeder cells. Some of them have already been elucidated, although much research is still needed to understand the biology of stem cell maintenance in culture. Traditionally, primary mouse embryonic fibroblasts (PMEFs) have been used as feeder layers, and both murine and human fibroblast cell lines have been shown to support pluripotency and self-renewal of hESC. Here we report the derivation of three new mouse embryonic fibroblast cell lines, MEFLU-M, MEFLU-T, and MEFLU-TB, with different properties regarding growth and support for undifferentiated hESCs. MEFLU-TB is able to support continuous growth of the newly derived Man-1, as well as H1, HUES-1, HUES-7, HUES-8, and HUES-9 human embryonic stem cell lines. After more than 50 passages and doublings, MEFLU-TB feeders compare to early passage primary mouse embryonic fibroblasts in their ability to support undifferentiated hESC growth. Our results contradict a previous paradigm that PMEFs tend to lose their capacity to support proliferation of hESCs with increasing passages, and show that the MEFLU-TB mouse embryonic fibroblast cell line and its conditioned medium have the potential to support the maintenance of hESC lines. Also, our results clearly show that spontaneous immortalization of primary fibroblasts can be achieved in culture without any chemical addition or genetic modification.

This study reports on a pediatric case of hemangiopericytoma (HPC) showing trisomy 15 as a sole anomaly. Trisomy 15 was observed in a total of 11 cells harvested at a very early passage from two different in-situ cultures. Trisomy 15, as a sole anomaly, has been described in hematologic disorders such as myelodysplastic syndromes but, to our knowledge, has never been documented in solid tumors. This is the first report of HPC with trisomy 15.

The culture medium supplemented with carp serum and fetal bovine serum (FBS) promoted cell growth significantly and induced morphological change of goldfish fin cells in early passage as compared to the medium containing FBS alone. However, these effects were not observed in RBCF-1, a cell line established from the goldfish fin. The sensitivity of the cells in early passage to carp serum suggests the following possibilities: (1) cells in early passage retain the ability to respond to growth-promoting factors specifically included in carp serum; and (2) this ability is lost during the process of long-term culture and/or long-term culture in FBS eliminates cell groups showing high dependency of cell growth on carp serum.

Two things happen every time an employee retires, is made redundant or leaves to join another company. The experience acquired at the company’s considerable expense literally walks out of the door and the company has to replace the individual, often from outside. For organizations, job change, which has always been continuous, has been accelerating over the past decade as individuals switch their employer every six years. Even though most professionally managed companies run induction courses, most new managers will readily admit that the components that most inhibit their early passage to full productivity relate to understanding and accommodating their new employer’s individual corporate culture, management and communication styles, and the detail of recent events. Because of the difficulties of imparting such information, companies generally leave individuals to assimilate these intangibles as best they can, usually by osmosis. The evidence suggests that it can take 12 months ‐ and often much longer ‐ to become fully productive. Lack of continuity has an insidious effect on productivity and competitiveness. Outlines a cost‐effective solution to high job mobility.

Biobanking became a necessity for translating genetic discoveries into clinical practice. Approaches to personalized medicine require a new model system for functional and pharmacogenomic studies of a variety of accumulating genetic variations, as well as new research environments such as biobankomics. Human lymphoblastoid cell lines (LCLs) will provide a valuable tool to meet such new demands in the biobankomics era. The National Biobank of Korea (NBK), which is leading the Korea Biobank Project, has a large collection of LCLs derived mostly from population-based cohort samples. Using a special long-term subculture collection of NBK LCLs, biological characteristics of early passage LCLs and terminally immortalized LCLs have been investigated to promote the utilization of LCLs and provide well quality-controlled LCLs for genetic and pharmacogenomic studies. As LCLs have been successfully phenotyped for cytotoxicity in response to various stimulators, including chemotherapeutic agents, environmental chemicals and irradiation, the utility of LCLs will increase in the future. Here, we discuss current and future applications of NBK LCLs for the biobankomics era.

A chromosome polymorphism was detected between two early passage euploid Chinese hamster cell strains when a fluorescence shift of the small metacentric No. 9 chromosome was resolved by flow cytometry. The characteristics of the polymorphism were studied using cultures established from ear clippings taken from 16 additional hamsters from our breeding colony. Additional variants of chromosome 9 were detected using flow cytometry, and a subset of these variants were analyzed by G- and C-banding. An increase of fluorescence recorded by flow cytometry correlated with an increase of centromeric heterochromatin. Autosomal normalization of the flow karyotype from 18 different animals indicated three distinct peak positions for chromosome 9. The results indicate that a discrete block of constitutive heterochromatin may be present in one or two extra copies within the small inbred colony of hamsters studied. To determine the inheritance patterns, hamsters with known polymorphic No. 9 chromosomes were bred. The flow karyotypes derived from the offspring of these matings provide strong evidence that chromosomal polymorphisms are inherited in Mendelian fashion.

The quantity and type of microsomal cytochrome P-450 was determined in early passage cultured fibroblasts from six mammalian species of widely differing life spans. The proportion of cytochrome P-450 and P448 varied considerably among species, with the shorter-lived species having a preponderance of cytochrome P-448. There was a good inverse correlation between potential species life span and the specific cytochrome P-448 content of cultured fibroblasts.

DNA replication of cultured fibroblasts of early passage derived from Werner's syndrome (adult progeria) patients and from normal subjects were compared by DNA fiber autoradiography. The frequency of replication initiation was decreased in Werner's syndrome cells derived from five patients compared with that in normal cells derived from three persons of different ages. The rate of DNA chain elongation did not differ between Werner's syndrome cells and normal cells.

Human cells in early passage cultures of benign oral tumors, normal oral tissues, normal lung tissues and, especially, in long-term established oral carcinoma cultures were highly susceptible to infection by human adenovirus types 5, 21, and 31. In contrast, replication of each adenovirus type was markedly limited in inoculated cells of newly-established oral squamous cell carcinoma cultures.

To estimate worker exposures to, and environmental contamination from, test chemicals and organic solvents used in an in vitro assay to assess the carcinogenic potential of chemicals, sodium fluorescein, a noncarcinogenic fluorescent material, was dissolved in tissue culture medium used to maintain early passage hamster embryo cells. Personal an environmental samples were taken over a 14-d period. The assay was performed according to standard procedures in a ventilated glove box or laminar flow safety cabinet. Considerably more than 99% of the chemical contamination found was recovered from the interiors of the glove box and hood and from disposable equipment. Contamination outside the containment units (less than 1 microgram) resulted from intralaboratory transport of chemicals, treated cultures, and contaminated equipment. We conclude that the standard operating particles and procedures provided adequate safeguards for personnel and the environment.

In the course of serial passages for several years a line of uncloned HeLa cells (A line) showed a gradual decrease in plaquing efficiency by coxsackievirus A 9 (CA 9 virus), while subcultures prepared from the same line kept frozen at an early passage level (A original line) did not show any change. However, it was observed later that the plaque-forming ability of the A original line (A orig. line) also decreased after serial passages as was observed with the A line. Comparing the characteristics of the same cell line at two different passage levels, it was found that the efficiency of adsorption of virus to cells was nearly the same, while virus yield at 8 hours after infection was different. The activity of alkaline phosphatase of cells was also different between these two passage levels, suggesting that a high enzymatic activity is associated with the susceptibility of cell cultures to CA 9 virus. Magnesium chloride at 25 mM enhanced plaque formation by CA 9 virus in highly passaged less susceptible cell cultures, and a possible role of the chemical as a stabilizer of alkaline phosphatase was discussed.

Isolation and propagation of bluetongue virus was attempted in 4 tissue culture systems. Good results were demonstrated in all 4 systems using high titre virus well adapted to tissue culture. Using material containing early passage virus of low titre no isolations were achieved in LK cells maintained in medium containing calf serum; the results were poor in LK cells with horse serum in the medium and in BHK cells with calf serum in the medium and only in BHK cells with horse serum incorporated in the medium were satisfactory results obtained. Substances inhibitory to BTV were demonstrated in varying amounts in 5 different batches of calf sera.