Reproducible methodology for the isolation of mesenchymal stem cells from human umbilical cord and its potential for cardiomyocyte generation

Abstract

Mesenchymal stem cells (MSCs) are considered to be a source of stem cells in tissue regeneration and therapeutics, due to their ability to undergo proliferation and differentiation. Complications associated with bone marrow-derived MSCs has prompted researchers to explore alternative sources of MSCs. The human umbilical cord is one such source; it is easily available and its collection is non-invasive. The sources of MSCs are non-controversial and thus they are not subjected to ethical constraints, as in the case of embryonic stem cells. MSCs are multipotent stem cells and has the ability to differentiate into various cell types of the mesodermal lineage. The aim of this study was to establish a reproducible method for the isolation of MSCs from human umbilical cord, as the few methods published till date gave inconsistent results and had a mixed population of contaminating endothelial cells. In our isolation strategy, we isolated a pure population of MSCs from Wharton's jelly of the human umbilical cord, which is very rich in collagen, and we used a high concentration of collagenase enzyme in the isolation of MSCs. Extensive phenotypic characterization analysis of these cells, using flow cytometry and antibody staining methods, have shown that we were able to isolate a pure population of the mesenchymal lineage cells that is devoid of haematopoietic and endothelial cell contaminants. When these MSCs were subjected to cardiomyocyte differentiation, we observed a change in the morphological characteristics, which was accompanied by the formation of myotube structures and spontaneous beating after 21 days.

Generation and characterization of human cardiac resident and non-resident mesenchymal stem cell

“…In contrast, hUC-MSCs retained a fibroblastic morphology until Cytotechnology ( 2016) 68:2061-2073 2069 passage 10 of the culture (data not shown). Although both MSCs were efficiently amplified under a standard culture condition (Pereira et al 2008;Hartmann et al 2010;Chong et al 2011), yet a prominent difference in term of doubling time and growth kinetics were noticed. Trypan blue and MTT assay clearly demonstrated that hUC-MSCs inherited a faster doubling time between 48 and 72 h than those of hC-MSC (*72 h).…”

“…Beside the targeted differentiation, MSCs also migrate and home at the injured/inflamed tissues to expedite the tissue repair by secreting growth factors and harnessing the uncontrolled immune response (Kinnaird et al 2004;Pereira et al 2008;Ramasamy et al 2008). Under a specific inductive condition, MSCs are capable of differentiating into mesodermal lineage cells, including cardiomyocytes (Sanchez et al 2000;Deans and Moseley 2000;Forbes et al 2002).…”

“…Although the surface antigen profiles and differentiation potential of hUC-MSC and hC-MSC are well documented, the electrophysiological properties of ion channels in undifferentiated hUC-MSCs and hC-MSC are still elusive (Shake et al 2002;Pereira et al 2008;He et al 2011;Bearzi et al 2009). Ion channels (delayed rectifier-like K ?…”

The success of stem cell transplantations and the potential post-transplantation complications may be dependent, among other factors, on the capacity of the recipient and the transplanted cells to repair DNA damage

“…They adhere to plastic surfaces in vitro, are capable of spontaneous differentiation along the osteogenic, chondrogenic and adipogenic lineages and express a subset (but not all) of the surface markers of the skin and lung fibroblasts (Dominici et al 2006). Targeted differentiation of MSCs may yield other cell types as well, such as insulin-producing pancreatic cells, different kidney-specific cell types such as tubulocytes and podocytes, cardiomyocytes, and other cell types , Pereira et al 2008, Brunner et al 2008, Asanuma et al 2010. MSCs possess significant potential for use in the regenerative and reparative medicine because of their versatility.…”

Ultrastructure of rat umbilical cord stroma-derived mesenchymal stem cells

“…Mesenchymal stem cells (MSCs) are self-renewing and multipotent cells, which have the potential to differentiate into various tissue cells, including osteoblasts (1,2), adipocytes, chondrocytes (3), neurons, glia cells (4), and cardiomyocytes (5), under favorable conditions. Owing to this potential, MSCs are being increasingly investigated for their use in cell-based regenerative medicine and tissue engineering (6).…”