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Co‐culture of mesenchymal‐like stromal cells derived from human foreskin permits long term propagation and differentiation of human embryonic stem cells
Author(s) -
Mamidi Murali Krishna,
Pal Rajarshi,
Mori Nor Azah Binti,
Arumugam Greetha,
Thrichelvam Saratha Thevi,
Noor Puteri J,
Abdullah Hj. Mohamad Farouk,
Gupta Pawan Kumar,
Das Anjan Kumar,
Zakaria Zubaidah,
Bhonde Ramesh
Publication year - 2011
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.23052
Subject(s) - mesenchymal stem cell , homeobox protein nanog , embryonic stem cell , biology , foreskin , microbiology and biotechnology , cd44 , cd90 , cd34 , stromal cell , cd146 , stem cell , induced pluripotent stem cell , cell culture , cancer research , cell , genetics , gene
Among the different parameters governing the successful derivation and expansion of human embryonic stem cells (hESC), feeder layers play the most important role. Human feeders in form of human mesenchymal stromal cells (hMSCs) and human foreskin fibroblasts (HFFs) lay the foundation for eradication of animal‐derived hESC culture system. In this study we explored the potential of human foreskin derived mesenchymal like stromal cells (HF‐MSCs) to support self renewal and pluripotency of hESC. The MSCs isolated from human foreskin were found to be resistant to standard concentrations and duration of mitomycin‐C treatment. Growth pattern, gene profiling (Oct‐4, Nanog, Sox‐2, Rex‐1), cytoskeletal protein expression (vimentin, nestin) and tri‐lineage differentiation potential into adipocytes, chondrocytes and osteocytes confirmed their mesenchymal stromal cell status. Further, the HF‐MSCs were positive for CD105, CD166, CD73, CD44, CD90, SSEA‐4, and negative for CD34, CD45, HLA‐DR cell‐surface markers and were found to exhibit BM‐MSC‐like characteristics. hESC lines co‐cultured with HF‐MSC feeders showed expression of expected pluripotent transcription factors Oct‐4, Nanog, Sox‐2, GDF‐3, Rex‐1, STELLAR, ABCG2, Dppa5, hTERT; surface markers SSEA‐4, TRA‐1‐81 and maintained their cytogenetic stability during long term passaging. These novel feeders also improved the formation of embryoid bodies (EBs) from hESC which produced cell types representing three germ layers. This culture system has the potential to aid the development of clinical‐grade hESCs for regenerative medicine and drug screening. Further, we envisage foreskin can serve as a valuable source of alternative MSCs for specific therapeutic applications. J. Cell. Biochem. 112: 1353–1363, 2011. © 2011 Wiley‐Liss, Inc.