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A Simple Method to Isolate and Expand Human Umbilical Cord Derived Mesenchymal Stem Cells: Using Explant Method and Umbilical Cord Blood Serum
International Journal of Stem Cells
Published online August 31, 2017;  
© 2017 Korean Society for Stem Cell Research.

Ghmkin Hassan1, Issam Kasem2,3, Chadi Soukkarieh2,3, Majd Aljamali1,3

1Department of Microbiology and Biochemistry, Faculty of Pharmacy, Damascus University, Damascus, Syria
2Department of Animal Biology, Faculty of Sciences, Damascus University, Damascus, Syria
3National Commission for Biotechnology (NCBT), Damascus, Syria
Correspondence to: Ghmkin Hassan
Department of Microbiology and Biochemistry, Faculty of Pharmacy, Damascus University, PO Box 10769, Damascus, Syria
Tel: +963112331919, Fax: +963112331919 E-mail: hsn.ghmkin@gmail.com
Co-Correspondence to Majd Aljamali
Department of Biochemistry and Microbiology, Faculty of Pharmacy, Damascus University, Damascus, Syria
Tel: +963115138306, Fax: +963115130104 E-mail: maljamali@gmail.com
; Accepted July 4, 2017.
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background and Objectives: Mesenchymal stem cells (MSCs) are multipotent stem cells that can be isolated from umbilical cords and are therapeutically used because of their ability to differentiate into various types of cells, in addition to their immunosuppressive and anti-inflammatory properties. Fetal bovine serum (FBS), considered as the standard additive when isolating and culturing MSCs, has a major limitation related to its animal origin. Here, we employed a simple and economically efficient protocol to isolate MSCs from human umbilical cord tissues without using digestive enzymes and replacing FBS with umbilical cord blood serum (CBS).
Methods and Results: MSCs were isolated by culturing umbilical cord pieces in CBS or FBS supplemented media. Expansion and proliferation kinetics of cells isolated by explant method in the presence of either FBS or CBS were measured, with morphology and multi-differentiation potential of expanded cells characterized by flow cytometry, RT-PCR, and immunofluorescence. MSCs maintained morphology, immunophenotyping, multi-differentiation potential, and self-renewal ability, with better proliferation rates for cells cultured in CBS compared to FBS supplement media.
Conclusions: We here present a simple, reliable and efficient method to isolate MSCs from umbilical cord tissues, where cells maintained proliferation, differentiation potential and immunophenotyping properties and could be efficiently expanded for clinical applications.
Keywords : Mesenchymal stem cells, Cord blood serum, Isolation, Differentiation


May 2017, 10 (1)