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Phenotypical and functional characteristics of mesenchymal stem cells from bone marrow: comparison of culture using different media supplemented with human platelet lysate or fetal bovine serum

机译:骨髓间充质干细胞的表型和功能特征:使用补充人血小板溶解产物或胎牛血清的不同培养基进行培养的比较

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Introduction Mesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several mesenchymal lineages, classically derived from bone marrow (BM) but potentially from umbilical cord blood (UCB). Although they are becoming a good tool for regenerative medicine, they usually need to be expanded in fetal bovine serum (FBS)-supplemented media. Human platelet lysate (HPL) has recently been proposed as substitute for safety reasons, but it is not yet clear how this supplement influences the properties of expanded MSCs. Methods In the present study, we compared the effect of various media combining autologous HPL with or without FBS on phenotypic, proliferative and functional (differentiation, cytokine secretion profile) characteristics of human BM-derived MSCs. Results Despite less expression of adipogenic and osteogenic markers, MSCs cultured in HPL-supplemented media fully differentiated along osteoblastic, adipogenic, chondrogenic and vascular smooth muscle lineages. The analyses of particular specific proteins expressed during osteogenic differentiation (calcium-sensing receptor (CaSR) and parathormone receptor (PTHR)) showed their decrease at D0 before any induction for MSC cultured with HPL mostly at high percentage (10%HPL). The cytokine dosage showed a clear increase of proliferation capacity and interleukin (IL)-6 and IL-8 secretion. Conclusions This study shows that MSCs can be expanded in media supplemented with HPL that can totally replace FBS. HPL-supplemented media not only preserves their phenotype as well as their differentiation capacity, but also shortens culture time by increasing their growth rate.
机译:简介间充质干细胞(MSCs)是能够分化为几种间充质谱系的多能细胞,其经典来源是骨髓(BM),但也有可能来自脐带血(UCB)。尽管它们已成为再生医学的良好工具,但通常需要在补充胎牛血清(FBS)的培养基中进行扩展。出于安全原因,最近有人提出了人血小板裂解液(HPL)的替代品,但目前尚不清楚这种补充剂如何影响扩增的MSC的特性。方法在本研究中,我们比较了自体HPL结合或不结合FBS的各种培养基对人BM来源的MSC的表型,增殖和功能(分化,细胞因子分泌特征)特征的影响。结果尽管脂肪形成和成骨标记物的表达较少,但在补充HPL的培养基中培养的MSC沿成骨细胞,脂肪形成,成软骨和血管平滑肌谱系完全分化。对成骨分化过程中表达的特定特定蛋白质(钙敏感受体(CaSR)和副激素受体(PTHR))的分析显示,在诱导诱导高密度脂蛋白(以高百分比(10%HPL)培养)的MSC之前,它们在D0时降低。细胞因子剂量显示明显增加的增殖能力以及白介素(IL)-6和IL-8分泌。结论这项研究表明,MSCs可以在补充了HPL的培养基中扩增,该培养基可以完全替代FBS。补充HPL的培养基不仅保留其表型和分化能力,而且通过提高其生长速度缩短培养时间。

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