Efficient generation of functional cardiomyocytes from human umbilical cord-derived virus-free induced pluripotent stem cells

Abstract

We have previously demonstrated that human umbilical cord-derived mesenchymal stem cells (UC-MSCs) can differentiate into cardiomyocyte-like cells. However, no contracting cells were observed during differentiation. In this study, we generated induced pluripotent stem cells (iPSCs) from UC-MSCs using mRNA reprogramming and focused on the differentiation of reprogrammed iPSCs into functional cardiomyocytes. For cardiac differentiation, the spontaneously contracting cell clusters were present on day 8 of differentiation. Immunostaining studies and cardiac-specific gene expression confirmed the cardiomyocyte phenotype of the differentiated cells. Electrophysiology studies indicated that iPSCs derived from UC-MSCs had a capacity for differentiation into nodal-, atrial-, and ventricular-like phenotypes based on action potential characteristics, and the derived cardiomyocytes exhibited responsiveness to β-adrenergic and muscarinic stimulations. Moreover, the derived cardiomyocytes displayed spontaneous intracellular Ca transients. These results demonstrate that functional cardiomyocytes can be generated from reprogrammed UC-MSCs, and the methodology described here will serve as a useful protocol to obtain functional cardiomyocytes from human mesenchymal stem cells.

Comparative analysis of mesenchymal stem/stromal cells derived from human induced pluripotent stem cells and the cognate umbilical cord mesenchymal stem/stromal cells

“…1 d; Supplement F. 1a-e). The above results indicated that human UMSCs could be successfully reprogrammed into iPSCs with well-defined cellular and molecular characteristic using the mRNA-based method which was in agreement with the previous study [ 8 , 9 , 32 ]. Overall, the above results strongly indicated that the iPSCs generated with the mRNA cocktails were human pluripotent stem cells with truly rejuvenation and pluripotent state and total two background iPSCs lines were established and used for further experiments in our study.…”

“…Also, another study has presented an optimized procedure to produce MSCs from human iPSCs depends on the generation of embryoid bodies (EBs) and transforming growth factor beta 1 (TGF-β1) [ 7 ]. In addition, Wu et al, has showed anti-BMP2 Ab/BMP2 immune complex was capable of promoting osteogenic differentiation of iPSCs going through iMSCs, which may be a favorable approach for iPSC-based bone tissue engineering [ 8 , 9 ]. Above studies have shown that iMSCs can be successfully differentiated from iPSCs with well-defined differentiation procedures.…”

Biological Characteristics of Umbilical Cord Mesenchymal Stem Cells and Its Therapeutic Potential for Hematological Disorders

“…Studies have shown that when body tissues suffer from ischemia-anoxia injury or chronic inflammation, the damaged tissue released chemokines, mobilized and guided the migration of MSCs to the injury site, and further induced differentiation into different types of cells ( Kidd et al, 2010 ). In vitro culture, UC-MSCs can be “trans-differentiated” under certain conditions to become mesoderm cells such as osteoblasts ( Xue et al, 2018 ) and cardiomyocytes ( Wu et al, 2018 ), endothelial cells ( Motawea et al, 2020 ), and also can differentiate into neurons in the ectoderm, hepatocytes ( Zhou et al, 2017 ), pancreatic cells ( Van Pham et al, 2014 ) in endoderm between germ layer. The low immunogenicity, high proliferation and differentiation potential of UC-MSCs make them seed cells for cell replacement therapy.…”

Human Umbilical Cord Mesenchymal Stem Cell Differentiation Into Odontoblast-Like Cells and Endothelial Cells: A Potential Cell Source for Dental Pulp Tissue Engineering

“…More importantly, compared to other sources of adult stem cells, there is a lower risk of viral contamination due to the protection provided by the placental barrier (Duya et al, 2013). In addition, studies have shown that hUCMSCs can differentiate into chondrocytes, osteoblasts, cardiomyocytes, and neurons, and hUCMSCs have been applied for cartilage regeneration, and bone tissue regeneration, and to treat kidney injury (Wang et al, 2016;Day et al, 2018;Wu et al, 2018;Zhang et al, 2018;Zheng et al, 2019). However, whether hUCMSCs can be applied in dental pulp regeneration remains unknown.…”