Overexpression of p21 Has Inhibitory Effect on Human Hematopoiesis by Blocking Generation of CD43＋ Cells via Cell-Cycle Regulation

Background and Objectives@#p21, an important member of the Cip/Kip family, is involved in inhibitory effects of RUNX1b overexpression during the early stage of human hematopoiesis. @*Methods@#and Results: We established a human embryonic stem cell (hESC) line with inducible expression of p21 (p21/hESCs). Overexpression of p21 did not influence either mesoderm induction or emergence of CD34＋ cells, but it significantly decreased the production of CD43＋ cells and changed the expression profile of hematopoiesis-related factors, leading to the negative effects of p21 on hematopoiesis. @*Conclusions@#In RUNX1b/hESC co-cultures when RUNX1b was induced from D0, perturbation of the cell cycle caused by upregulation of p21 probably prevented the appearance of CD43＋ cells, but not CD34＋ cells. The mechanisms via which CD34＋ cells are blocked by RUNX1b overexpression remain to be elucidated.

Quantitative evaluation of plasma after methylene blue and white light treatment in four Chinese blood centers.

BACKGROUND: Pathogen reduction technology is an important process in the blood safety system, including solvent/detergent treatment, filtration and methylene blue-photochemical technology (MB-PCT). To investigate the quality of MB-PCT-treated plasma, plasma samples from four Chinese blood centers were analyzed over 12 months of storage to determine the recovery of activities and levels of various plasma proteins. MATERIALS AND METHODS: Ten plasma units each from the Suzhou, Yancheng, Chongqing and Shandong blood centers were divided into two aliquots. One was subjected to treatment with one of two methylene blue-photochemical technology instruments and the other was used as control. The treated and untreated sample pairs were stored at -30°C. The recovery rates of coagulation factors, inhibitor proteins, total protein, immunoglobulins, and complement proteins were measured at different time points after storage. RESULTS: The mean recovery of most proteins exceeded 80% after MB treatment. The exceptions were factor XI and fibrinogen, of which only 71.3-74% and 69.0-92.3% were retained during storage. The two equipment types differed in terms of residual MB concentration in the plasma samples (0.18 µM and 0.29 µM, respectively). They had similar protein recovery rates at 0.5 month but differed at later time points. The four blood centers differed significantly with regard to factor II, V, VIII and fibrinogen activities. Only the samples from the Shandong blood center met the methylene blue treated fresh frozen plasma requirement. The major factor that influenced the quality of the MB-FFP samples was the time taken between blood collection and storage. DISCUSSION: Methylene blue treated plasma showed reduced coagulation factor (CF) activity and protein levels. The MB treatment-induced damage to the proteins was acceptable at the four Chinese blood centers, but the quality of the MB-treated plasma in general was not satisfactory. The main factor affecting plasma quality may be the duration of the collection and processing.

Three different bacterial detection systems for platelet concentrates under inter-laboratory conditions.

BACKGROUND: A variety of screening methods are currently used worldwide in order to decrease the risk of transfusion-transmitted sepsis and improve the safety of PCs. METHODS/MATERIALS: PCs inoculated with five different transfusion-relevant species of bacteria at concentrations of 1, 10, and 100 colony-forming units (CFU)ml(-1) were stored at 22°C for 7 days. Flow cytometry (FACS), BacT/Alert automated culturing, and a quantitative real-time PCR (Q-PCR) were then used to detect the presence of bacteria in samples prepared from these PCs. RESULTS: At the initial spiking concentrations of 1, 10, and 100 CFU ml(-1), Q-PCR detected all five bacterial species tested. Screening with the BacT/Alert culture-based system allowed bacterial detection (inoculated on day 0) within a mean time of 15.13 h for all three spiking concentrations. Using FACS, positive signals were obtained for all three concentrations of Escherichia coli and Bacillus cereus on day 1 and for initial spiking concentrations of Pseudomonas aeruginosa and Staphylococcus aureus of 1 CFU ml(-1) on day 2. For Staphylococcus epidermidis, detection of an initial inoculum of 1 CFU ml(-1) was possible only beginning on day 6. CONCLUSION: This study shows that under standard laboratory conditions the sensitivity of FACS in the detection of bacterial contamination of PCs was lower than that of either the BacT/Alert automated culturing method or Q-PCR.