In vitro differentiation of human skin-derived multipotent stromal cells into putative endothelial-like cells

Abstract

BackgroundMultipotent stem cells have been successfully isolated from various tissues and are currently utilized for tissue-engineering and cell-based therapies. Among the many sources, skin has recently emerged as an attractive source for multipotent cells because of its abundance. Recent literature showed that skin stromal cells (SSCs) possess mesoderm lineage differentiation potential; however, the endothelial differentiation and angiogenic potential of SSC remains elusive. In our study, SSCs were isolated from human neonatal foreskin (hNFSSCs) and adult dermal skin (hADSSCs) using explants cultures and were compared with bone marrow (hMSC-TERT) and adipose tissue-derived mesenchymal stem cells (hADMSCs) for their potential differentiation into osteoblasts, adipocytes, and endothelial cells.ResultsConcordant with previous studies, both MSCs and SSCs showed similar morphology, surface protein expression, and were able to differentiate into osteoblasts and adipocytes. Using an endothelial induction culture system combined with an in vitro matrigel angiogenesis assay, hNFSSCs and hADSSCs exhibited the highest tube-forming capability, which was similar to those formed by human umbilical vein endothelial cells (HUVEC), with hNFSSCs forming the most tightly packed, longest, and largest diameter tubules among the three cell types. CD146 was highly expressed on hNFSSCs and HUVEC followed by hADSSCs, and hMSC-TERT, while its expression was almost absent on hADMSCs. Similarly, higher vascular density (based on the expression of CD31, CD34, vWF, CD146 and SMA) was observed in neonatal skin, followed by adult dermal skin and adipose tissue. Thus, our preliminary data indicated a plausible relationship between vascular densities, and the expression of CD146 on multipotent cells derived from those tissues.ConclusionsOur data is the first to demonstrate that human dermal skin stromal cells can be differentiated into endothelial lineage. Hence, SSCs represents a novel source of stem/stromal cells for tissue regeneration and the vascularization of engineered tissues. Moreover, the CD146 investigations suggested that the microenvironmental niche might contribute to direct stromal cells multipotency toward certain lineages, which warrants further investigation.

Adult Stem Cell Subsets from Adult Human Dermis

“…Adult human dermal biopsies contain a variety of cells with discrete multipotent phenotypes [3][4][5][6][7][8][9][10][11], which are lost over extended culture periods [5]. The presence or absence of certain markers (CD73, CD90, CD105, CD146, CD271) associated with ex vivo multipotency appears to depend on factors such as biopsy procedure and expansion, cell density, culture conditions, and longevity of culture [5,9,10].…”

“…The presence or absence of certain markers (CD73, CD90, CD105, CD146, CD271) associated with ex vivo multipotency appears to depend on factors such as biopsy procedure and expansion, cell density, culture conditions, and longevity of culture [5,9,10]. LAVIV ® azficel-T cells contained 3% dermal derivatives expressing CD271 and 20% expressing CD146; positive and negative fractions for both phenotypes were independently collected.…”

“…Among these cellular surface markers, the canonical surface markers 5'-nucleotidase, ecto (CD73) or NT5E, thymocyte antigen 1 (CD90) or Thy-1 and Endoglin (CD105), were initially described as unique for mesenchymal stromal cells, but have since been identified as distributed among diverse types of human cells including fibroblasts with some or no in vitro mutipotency [1,8]. Previous research in human adult adipose cells and human neonatal dermal derivatives suggest Melanoma Cell Adhesion Molecule (CD146) or MCAM [10] and nerve growth factor receptor (CD271) or NGFR [11], respectively, as putative candidate markers to isolate mesenchymal stromal-like cells from human adult dermal derivatives, which would enhance the ability of forming bone, cartilage, and fat ex vivo. In theory, adult multipotent dermal fibroblasts can be easily attained through a minimally invasive and relatively painless skin punch biopsy [12] and hMSC derivatives purified via cell surface markers for in vitro or in vivo clinical applications [9,13].…”

Mesenchymal Stem Cells: A Future Option for Intervening Disease Management

“…The skin-derived stromal cells have shown the endothelial lineage differentiation in-vitro, and the angiogenic role with potential contribution to blood vessel formation in exvivo Chorioallantoic Membrane Assay (CAM) model is an excellent start for the pre-clinical considerations for the skin-derived MSCs. Therefore, human skin stromal cells are valuable resources that might be useful in applications requiring enhanced angiogenesis or in areas such as ischemic diseases [87][88][89]. Furthermore, these cells could be employed in tissue engineering and cell-based therapy in which vascularization is an essential component.…”

Evidence Supporting a Paracrine Effect of IGF-1/VEGF on Human Mesenchymal Stromal Cell Commitment

“…Periosteum-derived progenitor cells (PDPCs), which are located in the innermost layer of the periosteum, represent a promising cell source for bone tissue engineering strategies [Zhang et al, 2005[Zhang et al, , 2008Colnot et al, 2009;Chang and Knothe Tate, 2012;Ferretti et al, 2012] since the periosteum plays a critical role in endogenous repair processes [Colnot, 2009;Roberts et al, 2015]. Skin-derived MSCs (S-MSCs) could be another attractive stem cell population for skeletal tissue engineering approaches, as they are easily accessible without invasive procedures by means of skin biopsy [Takeda et al, 1992], and they have multipotent differentiation capability, being able to differentiate also toward an endothelial-like phenotype in a suitable culture system [Vishnubalaji et al, 2012].…”