In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells

Peter Szaraz; Yarden S. Gratch; Farwah Iqbal; Clifford L. Librach

doi:10.3791/55757

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Method Article

In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells

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DOI:

10.3791/55757

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August 9th, 2017

Peter Szaraz^*¹^,² , Yarden S. Gratch^*¹ , Farwah Iqbal¹^,² , Clifford L. Librach¹^,²^,³^,⁴^,⁵

¹Create Fertility Centre, ²Department of Physiology, University of Toronto, ³Department of Obstetrics and Gynecology, University of Toronto, ⁴Department of Physiology, University of Toronto, ⁵Department of Obstetrics and Gynecology, Women's College Hospital

^* These authors contributed equally

Summary

Here, we present a method to efficiently harness the cardiac differentiation potential of young sources of human mesenchymal stem cells in order to generate functional, contracting, cardiomyocyte-like cells in vitro.

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.

Introduction

Congestive heart failure (CHF) persists as a leading cause of morbidity and mortality worldwide. CHF often occurs following the massive loss of cardiomyocytes and the development of cell-free scar tissue as the pathological result of a myocardial infarction (MI)¹. While the heart is a partially self-renewing organ, the resident stem and progenitor cell pool responsible for executing tissue regeneration significantly diminishes in abundance and function in aged patients, often becoming insufficient for optimal recovery after injury. Thus, there is great interest in developing experimental treatments that involve the transplantation of healthy do....