Reprogramming of Human Umbilical Cord Stromal Mesenchymal Stem Cells for Myogenic Differentiation and Muscle Repair

Abstract

Human umbilical cord stromal mesenchymal stem cells (hUCS-MSCs) have the potential to differentiate into numerous cell types including epithelial cells, neurons and hepatocytes in vitro, in addition to mesenchyme-derived cells such as osteocytes, chondrocytes and adipocytes. One important property of these cells is the lack of type II major histocompatibility complex class molecules, thus allowing them to be considered as an excellent candidate for transplantations. Besides the use of 5-azacytidine as a supraphysiological inducer of myogenic transformation, no study has been published to date addressing the myogenic transformation efficiency of hUCS-MSCs by using a gene transfection strategy and/or co-culture with muscle cell lines. Here, we demonstrate the reprogramming efficiency of these cells, which differentiate into myocytes in vitro by MyoD transcription factor, the master regulator of skeletal muscle differentiation. Once induced via MyoD expression, hUCS-MSCs exhibited many cellular signs of myogenic conversion within 5 days and became capable of forming multinucleated myofibers, which exhibited all functional markers of fusion machinery such as β-catenin, neural cell adhesion molecule and M-cadherin as well as muscle cell-specific structural proteins including desmin, α-actinin, dystrophin, myosin heavy chain, and myoglobin together with muscle-specific enzyme, creatinine phosphokinase. Furthermore, programmed hUCS-MSCs were also capable of fusing with rat primary myoblasts to form heterokaryonic myotubes. Taken together, this study demonstrates the success of a novel cell reprogramming approach to be further evaluated at the in vivo level for use in restoring the defective dystrophin function as intrinsically found in the skeletal muscle fibers of Duchenne muscular dystrophy patients.

Fast-Degradable Microbeads Encapsulating Human Umbilical Cord Stem Cells in Alginate for Muscle Tissue Engineering

“…Only a few previous studies investigated myogenic differentiation of hUCMSCs by culturing the cells on dishes in vitro, or injecting the cells into animals. [14][15][16] None of these previous studies used any scaffold to seed hUCMSCs for myogenic differentiation. [14][15][16] In the present study, hUCMSC-encapsulating construct with fast-degradable microbeads in an AM was developed for muscle tissue engineering.…”

“…[14][15][16] None of these previous studies used any scaffold to seed hUCMSCs for myogenic differentiation. [14][15][16] In the present study, hUCMSC-encapsulating construct with fast-degradable microbeads in an AM was developed for muscle tissue engineering. The microbeads could degrade and release the cells throughout the AM, while creating macropores via microbead degradation.…”

“…[10][11][12][13] Only a few studies investigated hUCMSCs for muscle engineering by culturing on dishes, or injecting into animals. [14][15][16] To date, there has been no report on using a three-dimensional scaffold to seed hUCMSCs for myogenic differentiation.…”

Concise Review: Mesoangioblast and Mesenchymal Stem Cell Therapy for Muscular Dystrophy: Progress, Challenges, and Future Directions

“…Despite this, neither human nor canine ucMSCs expressed muscle proteins [38][39][40][41], indicating that ucMSCs lack myogenic potential in vivo in animal models for muscular dystrophy. However, forced expression of MyoD in ucMSCs enhanced skeletal muscle differentiation in vitro [42] and might also be useful to stimulate their myogenic differentiation in vivo, because this approach has been used to improve skeletal muscle differentiation of MSCs from synovium and orthopedic surgery remnants in vitro and in vivo [43,44].…”

Evaluation of adipose-derived stem cells for tissue-engineered muscle repair construct-mediated repair of a murine model of volumetric muscle loss injury

“…24,25 Stem cells offer an attractive alternative. To date, a host of stem cell types have been evaluated in muscle repair applications, including embryonic stem cells, 13,26 pluripotent adult stem cells, 22,27 muscle resident side population cells, 14 bone marrow-derived stem cells, 20,22 stromal cells isolated from synovial membrane, 21 human umbilical cord-derived cells, 28 pericytes, 15 and meso-angioblasts. 29 In this study, we evaluated the potential of adiposederived stem cells (ADSCs) as an alternative cell source for tissue-engineered skeletal muscle to circumvent the limited scaling ability of MDCs.…”