Sundaram Balasubramanian, Cherian Anne George, Kumar Sanjay
Center for Stem Cell Research, A Unit of inStem Bengaluru, Christian Medical College, CMC Rehab Campus Bagayam, Vellore, Tamil Nadu, 632002, India.
Department of Obstetrics and Gynaecology, Christian Medical College Vellore Hospital, Vellore, Tamil Nadu, 632002, India.
Methods Mol Biol. 2018;1577:35-53. doi: 10.1007/7651_2017_71.
Mesenchymal stem cells (MSCs) are derived from Wharton's jelly tissue of the human umbilical cord. Given appropriate culture conditions, these cells can self-renew and differentiate into multiple cell types across the lineages. Among the properties exhibited by these cells, immunomodulation through secretion of trophic factors has been widely exploited in a broad spectrum of preclinical/clinical regenerative applications. Moreover, the extracellular matrix is found to play a major role apart from niche cells in determining stem cell fate including that of MSCs. Therefore, the currently employed technique of two-dimensional culture expansion can alter the inherent properties of naïve MSCs originally residing within the three-dimensional space. This limitation can be overcome to some extent by using native extracellular matrix scaffold culture system which mimics the in situ microenvironment. In this chapter, we have elucidated the protocol for the preparation of a native extracellular matrix scaffold by decellularization of the MSC sheet and thereof culture expansion and characterization of human Wharton's jelly-derived MSCs.
间充质干细胞(MSCs)源自人脐带的华通氏胶组织。在适当的培养条件下,这些细胞可以自我更新并分化为跨谱系的多种细胞类型。在这些细胞表现出的特性中,通过分泌营养因子进行免疫调节已在广泛的临床前/临床再生应用中得到广泛应用。此外,发现细胞外基质除了在确定包括间充质干细胞在内的干细胞命运方面的小生境细胞外,还起着主要作用。因此,目前采用的二维培养扩增技术可以改变原本存在于三维空间中的原始间充质干细胞的固有特性。通过使用模拟原位微环境的天然细胞外基质支架培养系统,可以在一定程度上克服这一限制。在本章中,我们阐明了通过间充质干细胞片脱细胞制备天然细胞外基质支架及其培养扩增和人脐带华通氏胶来源的间充质干细胞表征的方案。