Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM
Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM
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DOI：
10.1080/14653240600920782
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发表时间：
2006-11-01
期刊：
影响因子：
4.5
通讯作者：
Dominici, M.
中科院分区：
文献类型：
--
作者：
Mueller, I.;Kordowich, S.;Dominici, M.
多能间充质基质细胞（MSC）已成为再生和移植医学的重要工具。越来越多的患者接受体外扩增的MSC。培养条件通常包括FSC，因为人血清不完全支持人MSC体外生长（MSCFCS）。对疯牛病、其他感染性并发症和宿主免疫反应的担忧推动了对替代培养补充剂的研究。方法由于血小板源生长因子长期以来被认为是MSC的生长因子，我们测试了补充血小板溶解产物的培养基以支持MSC增殖。结果我们发现BM来源的MSC的原代培养物可以用含有新鲜冷冻血浆和血小板的无动物血清培养基（MSCFFPP）建立。此外，MSCFFPP表现出旺盛的增殖，其上级优于含有FCS的经典培养条件。MSCFFPP的形态学与MSC FCS相当，MSCFFPP表达CD 73、CD 90、CD 105、CD 106、CD 146和HLA-ABC，而CD 34、CD 45和表面HLA-DR阴性，与预期一致。除了表型相同外，MSCFFPP还可以有效地分化为脂肪细胞和成骨细胞。在免疫调节特性方面，MSCFFPP与MSC FCS没有区别。MSC FFPP可抑制IL-2联合OKT-3或PHA诱导的PBMC增殖。讨论综上所述，在MSC培养中，出于临床目的，可以用FFPP安全地替代FCS。
Background Multipotent mesenchymal stromal cells ( MSC) have become important tools in regenerative and transplantation medicine. Rapidly increasing numbers of patients are receiving in vitro-expanded MSC. Culture conditions typically include FSC because human serum does not fully support growth of human MSC in vitro (MSCFCS). Concerns regarding BSE, other infectious complications and host immune reactions have fueled investigation of alternative culture supplements.Methods As PDGF has long been identified as a growth factor for MSC, we tested media supplementation with platelet lysate for support of MSC proliferation.Results We found that primary cultures of BM-derived MSC can be established with animal serum-free media containing fresh frozen plasma and platelets (MSCFFPP). Moreover, MSCFFPP showed vigorous proliferation that was superior to classical culture conditions containing FCS. MSCFFPP morphology was equivalent to MSC FCS, and MSCFFPP expressed CD73, CD90, CD105, CD106, CD146 and HLA-ABC while being negative for CD34, CD45 and surface HLA-DR, as expected. In addition to being phenotypically identical, MSCFFPP could efficiently differentiate into adipocytes and osteoblasts. In terms of immune regulatory properties, MSCFFPP were indistinguishable from MSC FCS. Proliferation of PBMC induced by IL-2 in combination with OKT-3 or by PHA was inhibited in the presence of MSCFFPP.Discussion Taken together, FCS can be replaced safely by FFPP in cultures of MSC for clinical purposes.