Brief Article
Copyright ©2013 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Stem Cells. Jan 26, 2013; 5(1): 26-33
Published online Jan 26, 2013. doi: 10.4252/wjsc.v5.i1.26
Fetal cardiac mesenchymal stem cells express embryonal markers and exhibit differentiation into cells of all three germ layers
Garikipati Venkata Naga Srikanth, Naresh Kumar Tripathy, Soniya Nityanand
Garikipati Venkata Naga Srikanth, Naresh Kumar Tripathy, Soniya Nityanand, Stem Cell Research Facility, Department of Hematology, Sanjay Gandhi Post-Graduate Institute of Medical Sciences, Lucknow 226014, India
Author contributions: Srikanth GVN designed and performed the experiments and analyzed the data; Tripathy NK and Nityanand S designed the study and analyzed the data; Nityanand S provided the reagents and analytical tools; Srikanth GVN, Tripathy NK and Nityanand S wrote the manuscript.
Supported by Department of Biotechnology, Government of India, BT/PR6519/MED/14/826/2005, to Dr. Nityanand S
Correspondence to: Soniya Nityanand, MD, PhD, FNASc, FASc, Professor, Head, Stem Cell Research Facility, Department of Hematology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Rae Bareli Road, Lucknow 226014, UP, India.
Telephone: +91-522-2494291 Fax: +91-522-2668017
Received: April 16, 2012
Revised: August 23, 2012
Accepted: December 20, 2012
Published online: January 26, 2013

AIM: To study the expression of embryonal markers by fetal cardiac mesenchymal stem cells (fC-MSC) and their differentiation into cells of all the germ layers.

METHODS: Ten independent cultures of rat fC-MSC were set up from cells derived from individual or pooled fetal hearts and studies given below were carried out at passages 3, 6, 15 and 21. The phenotypic markers CD29, CD31, CD34, CD45, CD73, CD90, CD105, CD166 and HLA-DR were analyzed by flow cytometry. The expression of embryonal markers Oct-4, Nanog, Sox-2, SSEA-1, SSEA-3, SSEA-4, TRA-1-60 and TRA 1-81 were studied by immunocytochemistry. The fC-MSC treated with specific induction medium were evaluated for their differentiation into (1) adipocytes and osteocytes (mesodermal cells) by Oil Red O and Alizarin Red staining, respectively, as well as by expression of lipoprotein lipase, PPARγ2 genes in adipocytes and osteopontin and RUNX2 genes in osteocytes by reverse-transcription polymerase chain reaction (RT-PCR); (2) neuronal (ectodermal) cells by expression of neuronal Filament-160 and Glial Fibrillar Acidic Protein by RT-PCR and immunocytochemistry; and (3) hepatocytic (endodermal) cells by expression of albumin by RT-PCR and immunocytochemistry, glycogen deposits by Periodic Acid Schiff staining and excretion of urea into the culture supernatant.

RESULTS: The fC-MSC expressed CD29, CD73, CD90, CD105, CD166 but lacked expression of CD31, CD34, CD45 and HLA-DR. They expressed embryonal markers, viz. Oct-4, Nanog, Sox-2, SSEA-1, SSEA-3, SSEA-4, TRA-1-81 but not TRA-1-60. On treatment with specific induction media, they differentiated into adipocytes and osteocytes, neuronal cells and hepatocytic cells.

CONCLUSION: Our results together suggest that fC-MSC are primitive stem cell types with a high degree of plasticity and, in addition to their suitability for cardiovascular regenerative therapy, they may have a wide spectrum of therapeutic applications in regenerative medicine.

Keywords: Fetal cardiac mesenchymal stem cells, Embryonal markers, Multipotent differentiation potential