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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۵۴-۵۴
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عنوان انگلیسی Ps-22: A Modified Stirred Tank Bioreactor for Self-Renewing Expansion of Hematopoietic Stem Cells
چکیده انگلیسی مقاله Objective: Performance evaluation of a modified stirred tank bioreactor (MSTB) having low shear stress for Ex vivo expansion of hematopoietic cell line. Materials and Methods: Medium was RPMI 1640 (Invitrogen/ 10% (v/v) fetal bovine serum (FBS, Gibco) with 1,00 U penicillin streptomycin/ml. U937 cell line (Invitrogen) as a model of hematopoietic stem cells (HSCs) was used to inoculate static and dynamic culture systems. To set up the MSTB system, cell proliferation place was separated from the stirred tank bioreactor (STB) and the system was under control by using a PLC system. Cell proliferation was evaluated by hemocytometer counting method. Cell morphology was also evaluated quantitatively by a parameter named cell roundness, Rc (Belview software). Cell surface marker of CD14 was also detected by flow cytometry (Becton- Dickinson) to test the cell self-renewal ability. Performance of the MSTB system was compared to the STB and T-flask cultures. Results: The U937 cell showed about 97.5 ± 1.7, 80 ± 2.8 and 0.9 ± 1.3 fold for the MSTB and T-flasks andSTB respectively. The almost all cells (93.7%) proliferated in the T-flasks during the 8-day culture period were CD14- cells. The percentage of CD14- cells, however, decreased to 73% and 11% in the cultures with MSTB and STB system at the same culture period. Interestingly, the frequencies of round-shaped cells (Rc > 0.9) for the T-flask, MSTB and STB cultures were 92.17% ± 2.1, 68% ± 4.9 and 18% ± 3.5 respectively. The MSTB system shows an acceptable performance as compared to the STB system due to providing lower shear stress on the cells. Conclusion: A modified stirred tank bioreactor system showed an acceptable performance for self-renewing expansion of hematopoietic cell line as HSC-mimicking cells in vitro, showing potential of the MSTB to possible use in the expansion HSCs in vitro.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/348
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