Human Alternatives to Fetal Bovine Serum for the Expansion of Mesenchymal Stromal Cells from Bone Marrow
Human Alternatives to Fetal Bovine Serum for the Expansion of Mesenchymal Stromal Cells from Bone Marrow
复制标题
DOI：
10.1002/stem.139
复制
发表时间：
2009-01-01
期刊：
影响因子：
5.2
通讯作者：
Klueter, Harald
中科院分区：
文献类型：
--
作者：
Bieback, Karen;Hecker, Andrea;Klueter, Harald
间充质基质细胞（MSC）是一种有前途的新型细胞治疗应用的候选人。对于临床规模生产，已建议将来自血清或血小板的人因子作为胎牛血清（FBS）的替代品。我们以前已经表明，合并的人血清（HS）和凝血酶激活的血小板释放血浆（tPRP）支持脂肪组织来源的MSC的扩增。骨髓（BM）来源的MSC的矛盾结果已经开始全面比较HS，tPRP和合并的人血小板裂解液（pHPL）和FBS对MSC分离，扩增，分化和免疫调节活性的影响。除了常规的Ficoll密度梯度离心外，使用谱系标志物表达细胞（RosetteSep）的耗竭和CD 271（+）分选用于BM-MSC富集。在含有10%FBS、HS、tPRP或pHPL的培养基中培养细胞。测定集落形成单位和累积群体倍增率，并最大限度地扩增MSC。虽然HS和tPRP都与FBS相当，支持分离和扩增，但pHPL显著加速BM-MSC增殖，在前两代内产生临床相关的数量。MSC的质量和功能，包括细胞表面标志物的表达，成脂和成骨分化，和免疫抑制作用是相似的MSC从所有的培养条件。重要的是，在任何培养条件下均未观察到自发细胞转化。在任何传代的培养物中均未检测到端粒酶活性。与先前来自脂肪组织来源的MSC的数据相反，发现pHPL是临床规模BM-MSC扩增中最合适的FBS替代物。干细胞2009; 27：2331-2341
Mesenchymal stromal cells (MSCs) are promising candidates for novel cell therapeutic applications. For clinical scale manufacturing, human factors from serum or platelets have been suggested as alternatives to fetal bovine serum (FBS). We have previously shown that pooled human serum (HS) and thrombin-activated platelet releasate in plasma (tPRP) support the expansion of adipose tissue-derived MSCs. Contradictory results with bone marrow (BM)-derived MSCs have initiated a comprehensive comparison of HS, tPRP, and pooled human platelet lysate (pHPL) and FBS in terms of their impact on MSC isolation, expansion, differentiation, and immunomodulatory activity. In addition to conventional Ficoll density gradient centrifugation, depletion of lineage marker expressing cells (RosetteSep) and CD271(+) sorting were used for BM-MSC enrichment. Cells were cultured in medium containing either 10% FBS, HS, tPRP, or pHPL. Colony-forming units and cumulative population doublings were determined, and MSCs were maximally expanded. Although both HS and tPRP comparable to FBS supported isolation and expansion, pHPL significantly accelerated BM-MSC proliferation to yield clinically relevant numbers within the first two passages. MSC quality and functionality including cell surface marker expression, adipogenic and osteogenic differentiation, and immunosuppressive action were similar in MSCs from all culture conditions. Importantly, spontaneous cell transformation was not observed in any of the culture conditions. Telomerase activity was not detected in any of the cultures at any passage. In contrast to previous data from adipose tissue-derived MSCs, pHPL was found to be the most suitable FBS substitute in clinical scale BM-MSC expansion. STEM CELLS 2009; 27: 2331-2341