<em>In Vitro</em> Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells

Abstract

Myocardial infarction and the subsequent ischemic cascade result in the extensive loss of cardiomyocytes, leading to congestive heart failure, the leading cause of mortality worldwide. Mesenchymal stem cells (MSCs) are a promising option for cell-based therapies to replace current, invasive techniques. MSCs can differentiate into mesenchymal lineages, including cardiac cell types, but complete differentiation into functional cells has not yet been achieved. Previous methods of differentiation were based on pharmacological agents or growth factors. However, more physiologically relevant strategies can also enable MSCs to undergo cardiomyogenic transformation. Here, we present a differentiation method using MSC aggregates on cardiomyocyte feeder layers to produce cardiomyocyte-like contracting cells. Human umbilical cord perivascular cells (HUCPVCs) have been shown to have a greater differentiation potential than commonly investigated MSC types, such as bone marrow MSCs (BMSCs). As an ontogenetically younger source, we investigated the cardiomyogenic potential of first-trimester (FTM) HUCPVCs compared to older sources. FTM HUCPVCs are a novel, rich source of MSCs that retain their in utero immunoprivileged properties when cultured in vitro. Using this differentiation protocol, FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to BMSCs, as indicated by the increased expression of cardiomyocyte markers (i.e., myocyte enhancer factor 2C, cardiac troponin T, heavy chain cardiac myosin, signal regulatory protein α, and connexin 43). They also maintained significantly lower immunogenicity, as demonstrated by their lower HLA-A expression and higher HLA-G expression. Applying aggregate-based differentiation, FTM HUCPVCs showed increased aggregate formation potential and generated contracting cells clusters within 1 week of co-culture on cardiac feeder layers, becoming the first MSC type to do so. Our results demonstrate that this differentiation strategy can effectively harness the cardiomyogenic potential of young MSCs, such as FTM HUCPVCs, and suggests that in vitro pre-differentiation could be a potential strategy to increase their regenerative efficacy in vivo.

Mesenchymal Stem Cell-Based Therapy for Cardiovascular Disease: Progress and Challenges

“…anti-inflammatory and antifibrotic effects; stimulate endogenous repair by paracrine signaling (growth factors and microvesicles); 32,87,88 limited capacity for direct tissue replacement [78][79][80] stimulate endogenous repair by paracrine signaling (growth factors and microvesicles); limited capacity for direct tissue replacement 41,48,178 unclear; evidence for direct remuscularization 24,172 or no evidence of remuscularization; 112 evidence for paracrine effects 37,112,169 modulates gene expression Molecular Therapy Vol. 26 No 7 July 2018 1611 www.moleculartherapy.org Review of hematopoietic lineage CD markers (CD45, CD34, CD14, CD19, and HLA-DR); be plastic adherent when maintained under standard culture conditions; and exhibit the ability to form into colony-forming unit fibroblasts (CFU-Fs) and differentiate into osteoblasts, adipocytes, and chondroblasts in vitro.…”

“…Under the proper conditions, MSCs are capable of transdifferentiating into functional cardiomyocytes in vivo and in vitro. [78][79][80] For example, transplantation of human MSCs, expressing a b-galactosidase reporter gene, into immunodeficient mice with experimental myocardial damage resulted in b-galactosidase + cardiomyocytes within the host myocardium, suggestive of cardiomyocyte differentiation of the xenografted MSCs. 81 Similarly, sex-mismatched cell therapy experiments, in porcine models of acute 32 and chronic MI, 82 provide evidence that engrafted male porcine MSCs differentiated into cardiomyocytes within the female host myocardium, as ascertained by co-localization of the Y chromosome with cardiac transcription factors GATA-4, Nkx2.5, and the structural cardiac protein a-sarcomeric actin.…”

Small G protein signaling modulator 3 (SGSM3) knockdown attenuates apoptosis and cardiogenic differentiation in rat mesenchymal stem cells exposed to hypoxia

“…In the present study, Sgsm3 KD ameliorated hypoxiainduced MSC death, which may be caused by Sgsm3 KD-induced increases in CX43. MI and subsequent ischemic processes under low oxygen conditions result in extensive cardiomyocyte loss, and MSC-based therapies are gaining attention as a way to replace current techniques [44]. Although complete differentiation into functional cells has not yet been achieved, MSCs can differentiate into cardiac cell types [10,44].…”

“…MI and subsequent ischemic processes under low oxygen conditions result in extensive cardiomyocyte loss, and MSC-based therapies are gaining attention as a way to replace current techniques [44]. Although complete differentiation into functional cells has not yet been achieved, MSCs can differentiate into cardiac cell types [10,44]. In vivo studies have demonstrated that MSC-derived differentiated cells have electrophysiological functions similar to cells originating from cardiac tissue [45,46].…”

Diferenciación de células madre derivadas de tejido adiposo (ADSCs) en cardiomiocitos a partir de co-cultivo indirecto :proliferación celular y expresión de proteínas cardíacas

“…La principal proteína de unión gap de comunicación eléctrica y bioquímica cardíaca es la conexina 43 (Cx43), además de estar presentes miosina de cadena pesada, alfa-actina y desmina (proteínas características de músculo estriado y músculo liso) [38].…”