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Background: The high incidence of septic transfusion reactions (STRs) led to testing being mandated by AABB from 2004. This was implemented by primary culture of single-donor apheresis platelets (APs) from 2004 and prestorage pooled platelets (PSPPs) from 2007.Study Design/methods: Platelet (PLT) aliquots were cultured at issue and transfusion reactions evaluated at our hospital. Bacterial contamination and STR rates (shown as rates per million transfusions in Results) were evaluated before and after introduction of primary culture by blood centers that used a microbial detection system (BacT/ALERT, bioMerieux) or enhanced bacterial detection system (eBDS, Haemonetics).Results: A total of 28,457 PLTs were cultured during pre-primary culture periods (44.7% APs; 55.3% at-issue pooled PLTs [AIPPs]) and 97,595 during post-primary culture periods (79.3% APs; 20.7% PSPPs). Forty-three contaminated units were identified in preculture and 34 in postculture periods (rates, 1511 vs. 348; p < 0.0001). Contamination rates of APs were significantly lower than AIPPs in the preculture (393 vs. 2415; p < 0.0001) but not postculture period compared to PSPPs (387 vs. 198; p = 0.9). STR rates (79 vs. 90; p = 0.98) were unchanged with APs but decreased considerably with pooled PLTs (826 vs. 50; p = 0.0006). Contamination (299 vs. 324; p = 0.84) and STR rates (25 vs. 116; p = 0.22) were similar for PLTs tested by BacT/ALERT and eBDS primary culture methods. A change in donor skin preparation method in 2012 was associated with decreased contamination and STR rates.Conclusion: Primary culture significantly reduced bacterial contamination and STR associated with pooled but not AP PLTs. Measures such as secondary testing near time of use or pathogen reduction are needed to further reduce STRs. [ABSTRACT FROM AUTHOR]

Aims The number of Sertoli cells during proliferative phase determines the fate of the germ cells in male reproductive system. A well-characterized cell line may help in better understanding of Sertoli cell biology. Hence, the present study assessed estradiol signaling in a mouse immature Sertoli cell line (MSC-1) as an alternative model in place of primary culture of Sertoli cells. Main methods In this study, we used MSC-1 cell line, derived from 10-day old mice. The cell cycle parameters were assessed, and the expression and regulation of Sertoli cell-specific secretory genes ( ABP ; androgen-binding protein ) and tight junction genes ( claudin-5 , occludin , and vimentin ) in response to estradiol was studied. Key findings The results obtained suggested the presence of both estrogen receptors (ERα and ERβ) in MSC-1 cells. In vitro scratch assay and cell-cycle analysis suggested the proliferative effects of estradiol in both time- and dose-dependent manner. The gene expression profiles of ABP , claudin-5 , and occludin showed biphasic regulation at low and high doses of estradiol. Analysis of signaling pathways suggested the activation of extracellular signal-regulated kinase (ERK) pathway with significantly increased pERK/ERK ratio ( p < 0.05). The results also suggested down regulation in the expression of mir-17 family members (mir-17, mir-20b, and mir-106a) ( p < 0.05). Significance Considering the limited number of Sertoli cell lines and long-term survival inability of primary culture of Sertoli cells, MSC-1 cells could be a potential cell line for understanding the mechanisms of various cellular events in Sertoli cells. [ABSTRACT FROM AUTHOR]

In this study, we report the enhanced osteoblast differentiation of mouse embryonic stem cell (mESC) on a novel polygalacturonic acid (PGA)/gelatin scaffold. The matrices of various PGA/geletin ratios were fabricated, biophysically characterized, and optimized for cell culture applications. Blended 2% PGA/gelatin scaffolds were highly porous and were robust with enhanced mechanical strength. Swelling studies showed they had high swelling capacity (six-fold higher than only gelatin scaffold) along with complete degradation in the presence of phosphate-buffered saline. Cytocompatibility of the matrices was evaluated using mouse fibroblast 3T3-L1 cell line showing normal spreading and proliferation as assessed by scanning electron microscopy (SEM) and MTT assay. Among different blends of PGA/gelatin, 2% PGA/gelatin (2PG) scaffold showed the optimum physical and biological compatibility for use in cell culture and differentiation of mESC, especially for osteoblast differentiation. The scaffold, as reported in this study, presents a promising tool for tissue engineering applications. [ABSTRACT FROM PUBLISHER]