Institute for Multiphase Processes, Leibniz Universität Hannover, Hannover, Germany.
Biochem Biophys Res Commun. 2011 Jul 29;411(2):317-22. doi: 10.1016/j.bbrc.2011.06.134. Epub 2011 Jun 25.
In regenerative medicine, human cell replacement therapy offers great potential, especially by cell types differentiated from immunologically and ethically unproblematic mesenchymal stem cells (MSCs). In terms of an appropriate carrier material, collagen scaffolds with homogeneous pore size of 65μm were optimal for cell seeding and cultivating. However, before clinical application and transplantation of MSC-derived cells in scaffolds, the safety and efficiency, but also possible interference in differentiation due to the material must be preclinically tested. The common marmoset monkey (Callithrix jacchus) is a preferable non-human primate animal model for this aim due to its genetic and physiological similarities to the human. Marmoset bone marrow-derived MSCs were successfully isolated, cultured and differentiated in suspension into adipogenic, osteogenic and chondrogenic lineages by defined factors. The differentiation capability could be determined by FACS. Specific marker genes for all three cell types could be detected by RT-PCR. Furthermore, MSCs seeded on collagen I scaffolds differentiated in adipogenic lineage showed after 28days of differentiation high cell viability and homogenous distribution on the material which was validated by calcein AM and EthD staining. As proof of adipogenic cells, the intracellular lipid vesicles in the cells were stained with Oil Red O. The generation of fat vacuoles was visibly extensive distinguishable and furthermore determined on the molecular level by expression of specific marker genes. The results of the study proved both the differential potential of marmoset MSCs in adipogenic, osteogenic and chondrogenic lineages and the suitability of collagen scaffolds as carrier material undisturbing differentiation of primate mesenchymal stem cells.
在再生医学中，人类细胞替代疗法具有巨大的潜力，尤其是通过分化自免疫和伦理上无问题的间充质干细胞（MSCs）的细胞类型。就合适的载体材料而言，孔径均一为 65μm 的胶原支架最适合细胞接种和培养。然而，在将 MSC 衍生细胞应用于支架进行临床应用和移植之前，必须对其安全性和效率进行临床前测试，同时还必须测试材料对分化的可能干扰。普通狨猴（Callithrix jacchus）由于其遗传和生理上与人类的相似性，是一种较好的非人类灵长类动物模型。已经成功分离、培养和悬浮分化狨猴骨髓来源的 MSC，使其分化为成脂、成骨和成软骨谱系，这是通过定义的因子实现的。通过 FACS 可以确定分化能力。通过 RT-PCR 可以检测到所有三种细胞类型的特异性标记基因。此外，接种在胶原 I 支架上的 MSC 在成脂谱系中分化 28 天后，细胞活力高且均匀分布在材料上，这通过 calcein AM 和 EthD 染色得到了验证。作为成脂细胞的证明，细胞内的脂质小泡用油红 O 染色。可以在细胞内明显区分和识别脂滴的形成，并通过特定标记基因的表达在分子水平上进一步确定。该研究结果证明了狨猴 MSC 在成脂、成骨和成软骨谱系中的分化潜力，以及胶原支架作为载体材料不干扰灵长类间充质干细胞分化的适用性。
