Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches: Further insights in the search for a fetal calf serum substitute
Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches: Further insights in the search for a fetal calf serum substitute
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DOI：
10.1002/jcp.20911
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发表时间：
2007-04-01
影响因子：
5.6
通讯作者：
Locatelli, F.
中科院分区：
文献类型：
--
作者：
Bernardo, M. E.;Avanzini, M. A.;Locatelli, F.
间充质基质细胞（MSCs）在细胞治疗和组织工程中的应用引起了人们极大的兴趣。目前，MSC在胎牛血清（FCS）的存在下在体外扩增;然而，FCS在临床级制剂中使用时引起了关注。本研究的目的是评估在补充有血小板裂解物（PL）的培养基中扩增的MSC是否具有适用于细胞治疗方法的生物学特性，该培养基已经显示出促进MSC生长。我们证实了先前发表的数据，显示在FCS或PL中扩增的MSC显示出相当的形态、表型和分化能力，而PL-MSC在克隆形成效率和增殖能力方面是上级的。我们进一步扩展了这些数据，通过研究免疫调节作用的骨髓间充质干细胞对同种异体抗原特异性免疫反应的混合淋巴细胞培养（MLC）。我们发现MSCs-PL在以下方面与MSCs-FCS相当：（i）降低同种异体抗原诱导的细胞毒性活性;（ii）有利于表达Treg表型的CD 4（+）T细胞亚群的分化;（iii）增加MLC上清液中IL-10的早期分泌，以及诱导IL-6产生的显著增加。与MSCs-PL相比，MSCs-FCS能更有效地抑制同种异体抗原诱导的淋巴细胞亚群增殖，减少IFN γ的早期分泌。通过分子核型分析和长期体外培养后正常形态/表型的维持证明了扩增的MSC对自发转化为肿瘤细胞的抗性。我们的数据支持MSC的免疫功能可塑性，并建议MSCs-PL可用作MSCs-FCS的替代品，尽管后者的细胞可能更适合于预防/治疗同种异体反应性相关的免疫并发症。
There is great interest in mesenchymal stromal cells (MSCs) for cell-therapy and tissue engineering approaches. MSCs are currently expanded in vitro in the presence of fetal calf serum (FCS); however, FCS raises concerns when used in clinical grade preparations. The aim of this study was to evaluate whether MSCs expanded in medium supplemented with platelet-lysate (PL), already shown to promote MSC growth, are endowed with biological properties appropriate for cell-therapy approaches. We confirm previously published data showing that MSCs expanded in either FCS or PL display comparable morphology, phenotype, and differentiation capacity, while PL-MSCs were superior in terms of clonogenic efficiency and proliferative capacity. We further extended these data by investigating the immune-regulatory effect of MSCs on the alloantigen-specific immune response in mixed lymphocyte culture (MLC). We found that MSCs-PL are comparable to MSCs-FCS in their capacity to: (i) decrease alloantigen-induced cytotoxic activity; (ii) favor differentiation of CD4(+) T-cell subsets expressing a Treg phenotype; (iii) increase early secretion of IL-10 in MLC supernatant, as well as induce a striking augmentation of IL-6 production. As compared with MSCs-PL, MSCs-FCS were more efficient in suppressing alloantigen-induced lymphocyte subset proliferation and reducing early IFN gamma-secretion. Resistance to spontaneous transformation into tumor cells of expanded MSCs was demonstrated by molecular karyotyping and maintenance of normal morphology/phenotype after prolonged in vitro culture. Our data support the immunological functional plasticity of MSCs and suggest that MSCs-PL can be used as an alternative to MSCs-FCS, although these latter cells might be more suitable for preventing/treating alloreactivity-related immune complications.