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Mesenchymal stem cell separation from Wharton’s jelly and its differentiation into keratinocytes

Mesenchymal stem cell separation from Wharton’s jelly and its differentiation into keratinocytes Mesenchymal stem cells (MSCs) are a group of cells present in bone marrow stroma, cord blood, Wharton’s jelly, and the stroma of various organs with the capacity for mesoderm-like cell differentiation. The aim of the present work is to isolate MSCs from human umbilical cord Wharton’s jelly and to examine the differentiation potential of these (MSCs) toward the epithelial lineage. Human MSCs derived from Wharton’s jelly of umbilical cord (≈31 cm) after a full-term delivery of 10 cases were localized and isolated and further cultured to enrich and enhance their proliferation. These MSCs were detected after an average of 21–30 days not only morphologically, as a uniform spindle fibroblast-like cells, and reached 70–80 % confluence with a good cellular yield, but also via their immunophenotypic analysis which showed positivity for CD29, CD90, CD105, and CD166 and negativity for CD34. All these cells showed 100 % negativity with CK19 to ensure the absence of any epithelial cells. Later, their differentiation potential was determined by culturing with conditioning medium that include bone morphogenetic protein-4, ascorbic acid, and human epithelium growth factor in a complete medium. These sets of conditions resulted in the expression of keratinocyte markers, namely pancytokeratin (mean value 92 %) and cytokeratin19 (mean value 74 %) using immunohistochemistry. In conclusion, these findings may have a significant impact on studies of early human epithelial differentiation, functional genomics, pharmacological testing, cell therapy, and tissue engineering by helping to eliminate worrying ethical and technical issues. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Comparative Clinical Pathology Springer Journals

Mesenchymal stem cell separation from Wharton’s jelly and its differentiation into keratinocytes

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Publisher
Springer Journals
Copyright
Copyright © 2013 by Springer-Verlag London
Subject
Medicine & Public Health; Pathology; Hematology; Oncology
eISSN
1618-565X
DOI
10.1007/s00580-013-1702-z
Publisher site
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Abstract

Mesenchymal stem cells (MSCs) are a group of cells present in bone marrow stroma, cord blood, Wharton’s jelly, and the stroma of various organs with the capacity for mesoderm-like cell differentiation. The aim of the present work is to isolate MSCs from human umbilical cord Wharton’s jelly and to examine the differentiation potential of these (MSCs) toward the epithelial lineage. Human MSCs derived from Wharton’s jelly of umbilical cord (≈31 cm) after a full-term delivery of 10 cases were localized and isolated and further cultured to enrich and enhance their proliferation. These MSCs were detected after an average of 21–30 days not only morphologically, as a uniform spindle fibroblast-like cells, and reached 70–80 % confluence with a good cellular yield, but also via their immunophenotypic analysis which showed positivity for CD29, CD90, CD105, and CD166 and negativity for CD34. All these cells showed 100 % negativity with CK19 to ensure the absence of any epithelial cells. Later, their differentiation potential was determined by culturing with conditioning medium that include bone morphogenetic protein-4, ascorbic acid, and human epithelium growth factor in a complete medium. These sets of conditions resulted in the expression of keratinocyte markers, namely pancytokeratin (mean value 92 %) and cytokeratin19 (mean value 74 %) using immunohistochemistry. In conclusion, these findings may have a significant impact on studies of early human epithelial differentiation, functional genomics, pharmacological testing, cell therapy, and tissue engineering by helping to eliminate worrying ethical and technical issues.

Journal

Comparative Clinical Pathology – Springer Journals

Published: Feb 24, 2013

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