Biological Features of Human Bone Marrow Stromal Cells (hBMSC) Cultured with Animal Protein-Free Medium-Safety and Efficacy of Clinical Use for Neurotransplantation
Biological Features of Human Bone Marrow Stromal Cells (hBMSC) Cultured with Animal Protein-Free Medium-Safety and Efficacy of Clinical Use for Neurotransplantation
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DOI：
10.1007/s12975-011-0088-y
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发表时间：
2011-09-01
影响因子：
6.9
通讯作者：
Houkin, Kiyohiro
中科院分区：
文献类型：
--
作者：
Shichinohe, Hideo;Kuroda, Satoshi;Houkin, Kiyohiro
供体细胞在无血清培养基中的培养对于细胞移植治疗的临床应用具有重要意义。本研究的目的是验证一个假设，即人骨髓基质细胞（hBMSC）扩增胎牛血清（FCS），含血小板裂解液（PL）的培养基保留其生物学特性，有利于中枢神经系统的再生。hBMSC用5%PL或10%FCS培养。用流式细胞术分析其表型，用酶联免疫吸附测定法定量其生长因子的产生。免疫细胞化学染色证实其具有神经分化能力。hBMSC-FCS和hBMSC-PL在形态学和细胞表面标志方面无明显差异。两例患者均为CD 44、CD 90、CD 105和CD 166阳性，CD 34、CD 45和CD 271阴性。人脑源性神经营养因子、人肝细胞生长因子、人β-神经生长因子和人血小板源性生长因子-BB的产生在两组之间没有差异，尽管hBMSC-PL产生的TGF-β 1的量显著高于hBMSC-FCS。两组细胞在体外向神经元和星形胶质细胞的分化无显著差异。用含PL的培养基扩增的hBMSC保留了其神经分化和神经保护的生物学能力。PL在扩增hBMSC用于细胞治疗方面可能是FCS的一种有临床价值和安全的替代物。
The donor cell culture in animal serum-free medium is quite important for the clinical application of cell transplantation therapy. This study was aimed to test the hypothesis that the human bone marrow stromal cells (hBMSC) expanded with fetal calf serum (FCS)-free, platelet lysate (PL)-containing medium retain their biological features favoring central nervous system regeneration. The hBMSC were cultured with 5% PL or 10% FCS. Their phenotypes were analyzed with flow cytometry, and their production of growth factors was quantified with enzyme-linked immunosorbent assay. Their capacity of neural differentiation was verified by immunocytochemistry. There was no significant difference in morphology and cell surface marker between the hBMSC-FCS and hBMSC-PL. Both of them were positive for CD44, CD90, CD105, and CD166 and were negative for CD34, CD45, and CD271. The production of human brain-derived neurotrophic factor, human hepatocyte growth factor, human beta-nerve growth factor, and human platelet-derived growth factor-BB did not differ between the two groups, although the hBMSC-PL produced significantly more amount of TGF-beta 1 than the hBMSC-FCS. There was no significant difference in their in vitro differentiation into the neurons and astrocytes between the two groups. The hBMSC expanded with PL-containing medium retain their biological capacity of neural differentiation and neuroprotection. The PL may be a clinically valuable and safe substitute for FCS in expanding the hBMSC for cell therapy.