Direct differentiation of cord blood derived mesenchymal stem cells into keratinocytes without feeder layers and cAMP inducers

Abstract

Objectives: The purpose of our study was isolation of umbilical cord blood derived mesenchymal stem (UCB-MSCs), their direct differentiation towards keratinocytes without using feeder layers, cAMP inducers and hormones known for morphological maintenance and proliferation of keratinocytes and characterization of UCB-MSCs through flowcytometry and keratinocytes through immunofluorescence.
 Methods: We have isolated and cultured UCB-MSCs (n=4) following critical parameters for successful isolation like sample processing within an hour of collection, gestational age not more than 38 weeks, no co-morbid and blood volume at least 80 ml. Cord blood mononuclear cells were isolated through ficoll based density-gradient centrifugation then cultured to isolate MSCs, defined by minimum criteria of International Society for Cellular Therapy. UCB-MSCs were then differentiated directly into keratinocytes. Differentiation was confirmed by morphology and characterized through immunofluorescence staining. UCB samples were collected from gynae/obstetric ward of OJHA campus under sterile conditions and processed at Stem cells and Regenerative medicine Lab, Dow Research Institute of Biotechnology and Biomedical Sciences, Ojha campus. The total duration of study was approximately 12 months.
 Results: We have successfully isolated UCB-MSCs that were plastic adherent, spindle shaped, showed trilineage mesodermal differentiation potential and were positive for CD90, CD73 and CD105 and negative for CD34 markers. UCB-MSCs were directly differentiated towards keratinocytes without using cAMP inducers, hormones or feeder layers. Differentiated keratinocytes attained typical honeycomb morphology and were stained positive on immunofluorescence for anti-pan cytokeratin antibody.
 Conclusion: Our study concludes possibility of direct differentiation of isolated and cultured UCB-MSCs into keratinocytes without using feeder layers and conventional keratinocyte culture media.
 doi: https://doi.org/10.12669/pjms.36.5.1566
 How to cite this:Ghauri AK, Wahid M, Mirza T, Uddin JAA. Direct differentiation of cord blood derived mesenchymal stem cells into keratinocytes without feeder layers and cAMP inducers. Pak J Med Sci. 2020;36(5):---------. doi: https://doi.org/10.12669/pjms.36.5.1566
 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

The Creation of Human Epithelial Tissue Graft Derived From Human Blood Cord Mesenchymal Stem Cells and Human Amniotic Membrane to Be Applied in Emergency Care

“…The cells in this human like-epithelial tissue express characteristic features very close to skin cells epithelium, such as the markers p63 and ck 1/10 (Fi. [1][2][3][4][5][6][7][8]. The combined use of hUCB-MSCs and hAM presents numerous advantages from both ethical and purely clinical point of view.…”

“…Therefore, the need to develop a solution to create autologous skin tissue has become strong. At the moment, there are many products used to support the skin healing process of both biological and chemical origin [5][6][7][8][9][10]. However, the common disadvantage of these products is that they do not contain live cells or lack all the factors and molecules necessary for real and effective integration with the host's natural skin tissue [10][11][12][13][14][15].…”

“…Human UCB contains pluripotent and multipotent MSCs able of differentiating into many different cell phenotypes [5,9]. These cells are also capable to differentiate into epithelial-like cells, an excellent advantage for creating epithelial surrogate for tissue bio-engineering with a basic structure similar to biological skin tissue [7,[27][28][29][30][31].…”