Umbilical Cord-Derived Mesenchymal Stem Cells Isolated by a Novel Explantation Technique Can Differentiate into Functional Endothelial Cells and Promote Revascularization

Abstract

Stem cells transplantation holds great promise for the treatment of ischemic diseases through functional revascularization. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are also an ideal candidate for cell-based bioengineering. Herein, we report on the development of a simple and effective protocol to isolate UC-MSCs, and confirm their endothelial potential both in vitro and in vivo. UC-MSCs were isolated by a novel explantation technique and induced to differentiate into endothelial-like cells. Then UC-MSCs were transplanted into ischemic mouse model and cultured on 3D gel/MMT-CS composite scaffolds. Morphological and proliferation assessments show that sufficient UC-MSCs can be generated during a relatively short culture period with explantation technique. Increased expression of endothelial-specific markers (KDR and vWF), and functional markers (ac-LDL uptake and UEA-1 binding), indicate that functional endothelial progenitor cells are induced after 9 days of in vitro culture. In an ischemic hindlimb mouse model, the ratio of ischemic/nonischemic limb perfusion 4 weeks after MSCs transplantation reached 0.84 +/- 0.09. The capillary density of this group was 2.57-fold greater than that of sham-injected mice (P < 0.05). Immunofluorescence and immunohistological analyses indicate that MSCs may act to salvage the ischemic tissue by incorporating into the local vasculature. In vitro, UC-MSCs were observed to incorporate into 3D gel/MMT-CS composite scaffolds, to secrete extracellular matrix, to remain viable, and to retain their proliferation capacity. In conclusion, UC-MSCs isolated by novel yet simple explantation technique are well suited for application in the development of novel stem cell-based revascularization therapies.

Mesenchymal stem cells derived from Wharton jelly of the human umbilical cord ameliorate damage to human endometrial stromal cells

“…In the second step, we used the 10, 20, 40, 60, 80, 100, 1,000 mM concentrations of mifepristone treatment with ESCs for 48 hours, then detected the IC of withdrawa l of mifepristone after 24, 48, 72, and 96 hours. The result indicated that IC50 value (16,21) in both phase ESCs was $60 mmol/L with withdrawal of mifepristone after 48 hours ( Fig. 1E and F).…”

“…WJ-MSCs were isolated and cultured by adopting the explantation technique as described by Xu et al (16). These human UC-MSCs had a typical spindle-shaped appearance and were positive for CD 73 (97.3 AE 2.1%), 90 (98.9 AE 0.9%), and 105 (97.4 AE 1.6%) and negative for CD 14 (0.2 AE 0.08%), 34 (0.9 AE 0.05%), 45 (0.4 AE 0.03%), and 79a (0.1 AE 0.03%) and HLA-DR (0.5 AE 0.2%) (CD14-FITC, CD45-FITC, CD79a-APCCD90-APC, CD34-PE, CD73-PECD105-PE, HLA-DR-PE.BD) by flow cytometric analysis.…”

Transplantation of umbilical cord–derived mesenchymal stem cells as a novel strategy to protect the central nervous system: technical aspects, preclinical studies, and clinical perspectives

“…Recently, a new method has been proposed for the isolation of MSCs from Wharton's jelly by the direct delivery of enzymatic solutions into the tissue, similar to the method used for the enzymatic digestion of the pancreas for Langerhans islet isolation (50). To avoid the disadvantages of enzymatic digestion, which has been shown to potentially alter cell proliferation and function, some groups have developed UC explant culture approaches that have been shown to be simple, reproducible, and efficient (63)(64)(65)(66). However, considering the heterogeneity of the processes used for cell isolation and expansion, and in view of their use for clinical applications, some groups have tried to develop optimized and standardized methods based on the use of xeno-and serum-free culture media (52,67,68) or by taking advantage of the plastic-adherence properties of MSCs that allow cell expansion without UC dissection or enzymatic digestion (69).…”

A Mathematical Model Predicting the Coculture Dynamics of Endothelial and Mesenchymal Stem Cells for Tissue Regeneration

“…MSCs cocultured with ECs were found to exhibit strong pro-angiogenic and vasculogenic effects that were associated with their ability to stabilize the formation of tubular vascular-like structures both in vitro 9,11 and in vivo. 12,13 MSCs were also shown, under certain in-vitro conditions, to trans-differentiate into ECs, [14][15][16] further reaffirming their in-vivo association. However, despite the ample literature reporting EC and stem cell cocultures, 3,5,17,18 no comprehensive investigation has explored and quantified their population dynamics, let alone investigated them together in a unifying model addressing the several factors influencing cell growth.…”