3D Decellularized Native Extracellular Matrix Scaffold for In Vitro Culture Expansion of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells (hWJ MSCs)
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.
KeywordsDecellularization Extracellular matrix Mesenchymal stem cells Scaffold Wharton’s jelly Stem cell niche
Alpha modification of minimum essential medium
Cluster of differentiation
Fluorescence-activated cell sorting
- hWJ MSCs
Human Wharton’s jelly-derived MSCs
Mesenchymal stem cells
Phosphate buffered saline
Phosphate buffered saline with Tween 20
We would like to thank the University Grants Commission (UGC) for granting Junior Research Fellowship (JRF)/Senior Research Fellowship (SRF) to Balasubramanian Sundaram. We would like to thank the Department of Biotechnology (DBT), India, for granting Ramalingaswami Fellowship and research support grant to Sanjay Kumar. We would like to thank Merylin Nixon, Department of Community Health, Christian Medical College Vellore Hospital, Vellore, India, for assisting us in obtaining umbilical cord tissue samples. We would like to appreciate the technical assistance provided by core facilities of Centre for Stem Cell Research, A unit of inStem Bengaluru, Christian Medical College, Vellore, Bagayam, Tamil Nadu, 632002, India.
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