Laboratory of Hematopoiesis, UPRES-EA3855, Faculty of Medicine, University François Rabelais, 10 boulevard Tonnellé, 37032 Tours Cedex, France.
Stem Cell Res Ther. 2012 Feb 14;3(1):6. doi: 10.1186/scrt97.
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.
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.
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.
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.
间充质干细胞(MSCs)是多能细胞,能够分化为几种间充质谱系,经典的来源于骨髓(BM),但潜在的来源于脐带血(UCB)。虽然它们正成为再生医学的良好工具,但通常需要在含有胎牛血清(FBS)的培养基中进行扩增。人血小板裂解液(HPL)最近因其安全性而被提议作为替代品,但目前尚不清楚这种补充物如何影响扩增的 MSCs 的特性。
在本研究中,我们比较了各种结合自体 HPL 与或不与 FBS 的培养基对人 BM 来源的 MSCs 的表型、增殖和功能(分化、细胞因子分泌谱)特性的影响。
尽管脂肪生成和成骨标记物的表达较少,但在 HPL 补充培养基中培养的 MSCs 可沿着成骨细胞、脂肪细胞、软骨细胞和血管平滑肌谱系完全分化。在成骨分化过程中表达的特定蛋白质(钙敏感受体(CaSR)和甲状旁腺素受体(PTHR))的分析表明,在诱导之前,在用 HPL 培养的 MSC 中,它们在 D0 时的表达减少,尤其是在高百分比(10%HPL)时。细胞因子剂量显示出增殖能力和白细胞介素(IL)-6 和 IL-8 分泌的明显增加。
本研究表明,MSCs 可以在含有 HPL 的培养基中扩增,HPL 可以完全替代 FBS。HPL 补充培养基不仅可以保留其表型和分化能力,还可以通过增加其生长速度来缩短培养时间。