Genetic factors influencing murine hematopoietic productivity in culture.
Genetic factors influencing murine hematopoietic productivity in culture.
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影响培养中小鼠造血生产力的遗传因素。
DOI：
10.1002/jcp.1041640113
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
VanZant,G
中科院分区：
文献类型：
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作者：
Phillips,RL;Couzens,MS;VanZant,G
为了研究之前描述的特定小鼠品系干细胞动力学中表现的遗传差异，我们测量了该推定基因 stk 对离体干细胞和祖细胞群生长和扩增的影响。将来自两种近交小鼠品系 C57BL/6J 和 DBA/2J 的骨髓细胞放入单独的生物反应器培养物中，连续灌注含有促红细胞生成素 (Epo)、白介素-3 (IL-3)、粒细胞巨噬细胞集落刺激因子 (GM-CSF) 和 Kit 配体以及 5% CO2 的生长培养基。在培养约 1 周内，DBA/2J 骨髓的细胞数量扩增至 20 倍，C57BL/6J 骨髓的细胞数量扩增至 10 倍。在细胞产生峰值之前，还观察到集落形成单位 (CFU)-GM 的大量产生（比输入水平增加九倍），而且重要的是，第 12 天集落形成单位脾脏 (CFU-S；两到三倍) 也发生了适度扩张，尽管 CFU-S 产量在相对较短的 4 天达到峰值。由于不利的生长条件或终末分化，培养物中的 CFU-S 和 CFU-GM 水平迅速下降。通过增加培养基灌注速率来去除有毒代谢物的尝试导致 DBA/2J 骨髓的细胞扩增能力增强。在骨髓接种前建立基质细胞的生物反应器中，CFU-S（尤其是 DBA/2J）和 CFU-GM 的扩增更大，尽管总细胞产量似乎不受影响，这可能是因为已经达到了最大细胞密度。 DBA/2J 骨髓中测量到的 CFU-S 扩增潜力相对于 C57BL/6J 的相对较高，这将有助于追踪遗传对骨髓生产能力的贡献。 © 1995 Wiley-Liss, Inc.
In order to study a previously described genetic difference manifested in stem cell kinetics of specific mouse strains, effects of this putative gene,stk, were measured on growth and expansion of stem and progenitor cell populations ex vivo. Bone marrow cells from each of two inbred mouse strains, C57BL/6J and DBA/2J, were placed into separate bioreactor cultures perfused continuously with growth medium containing erythropoietin (Epo), interleukin‐3 (IL‐3), granulocyte‐macrphage colony stimulating factor (GM‐CSF), and Kit ligand as well as 5% CO2. Expansion of cell numbers reached 20‐fold for DBA/2J and 10‐fold for C57BL/6J marrow within about 1 week of culture. Significant production was also seen of colonyforming unit (CFU)‐GM (up nine‐fold from input levels) just prior to the cell production peak, and, importantly, moderate expansion of day 12 colony‐forming unit‐spleen (CFU‐S; two‐ to threefold) occurred as well, although CFU‐S production peaked at a relatively short 4 days. CFU‐S and CFU‐GM levels declined rapidly in culture, either because of unfavorable growth conditions or terminal differentiation. Attempts to remove toxic metabolites by increasing the media perfusion rate resulted in a boost in cell expansion capability by DBA/2J marrow. In bioreactors in which stromal cells were established before marrow inoculation, there was greater expansion of CFU‐S (especially by DBA/2J) and CFU‐GM, although total cell yield appeared to be unaffected, perhaps because the maximum cell density had already been reached. The relative high potential for CFU‐S expansion measured in DBA/2J marrow over that of C57BL/6J will be useful in following genetic contributions to bone marrow production capacity. © 1995 Wiley‐Liss, Inc.