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3D Decellularized Native Extracellular Matrix Scaffold for In Vitro Culture Expansion of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells (hWJ MSCs)

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Decellularized Scaffolds and Organogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1577))

Abstract

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.

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Abbreviations

αMEM:

Alpha modification of minimum essential medium

APC:

Allophycocyanin

CD:

Cluster of differentiation

DAPI:

4′,6-Diamidino-2-phenylindole

DMSO:

Dimethyl sulfoxide

DNase:

Deoxyribonuclease

DPX:

Distyrene-plasticizer-xylene

ECM:

Extracellular matrix

EDTA:

Ethylenediaminetetraacetic acid

FACS:

Fluorescence-activated cell sorting

FITC:

Fluorescein isothiocyanate

hWJ MSCs:

Human Wharton’s jelly-derived MSCs

IgG:

Immunoglobulin G

ITS:

Insulin-transferrin-selenium

MSCs:

Mesenchymal stem cells

μl:

Microliters

μM:

Micromoles

ml:

Milliliters

mM:

Millimoles

PBS:

Phosphate buffered saline

PBST:

Phosphate buffered saline with Tween 20

PE:

Phycoerythrin

Pen-Strep:

Penicillin-Streptomycin

2D:

Two dimensional

3D:

Three dimensional

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Acknowledgments

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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Authors and Affiliations

  1. Center for Stem Cell Research, A Unit of inStem Bengaluru, Christian Medical College, CMC Rehab Campus Bagayam, Vellore, Tamil Nadu, 632002, India

    Balasubramanian Sundaram & Sanjay Kumar

  2. Department of Obstetrics and Gynaecology, Christian Medical College Vellore Hospital, Vellore, Tamil Nadu, 632002, India

    Anne George Cherian

  3. Department of Community Health, Christian Medical College Vellore Hospital, Vellore, Tamil Nadu, 632002, India

    Anne George Cherian

Authors
  1. Balasubramanian Sundaram
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  2. Anne George Cherian
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  3. Sanjay Kumar
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Corresponding author

Correspondence to Sanjay Kumar .

Editor information

Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, ON, Canada

    Kursad Turksen

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Sundaram, B., Cherian, A.G., Kumar, S. (2017). 3D Decellularized Native Extracellular Matrix Scaffold for In Vitro Culture Expansion of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells (hWJ MSCs). In: Turksen, K. (eds) Decellularized Scaffolds and Organogenesis. Methods in Molecular Biology, vol 1577. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_71

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  • DOI: https://doi.org/10.1007/7651_2017_71

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7655-3

  • Online ISBN: 978-1-4939-7656-0

  • eBook Packages: Springer Protocols

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