Hypoxia preconditioned bone marrow-derived mesenchymal stromal/stem cells enhance myoblast fusion and skeletal muscle regeneration

Karolina Archacka,Ilona Kalaszczyńska,Mirosław Janowski,Alicja Górzyńska,Gabriela Muras,Katarzyna Jańczyk-Ilach,Joanna Graffstein,Władysława Stremińska,Anna M Różycka,Piotr Walczak,Bartosz Mierzejewski,Marta Krawczyk,Edyta Brzoska,Iwona Grabowska,Maria A Ciemerych

doi:10.1186/s13287-021-02530-3

Abstract

BackgroundThe skeletal muscle reconstruction occurs thanks to unipotent stem cells, i.e., satellite cells. The satellite cells remain quiescent and localized between myofiber sarcolemma and basal lamina. They are activated in response to muscle injury, proliferate, differentiate into myoblasts, and recreate myofibers. The stem and progenitor cells support skeletal muscle regeneration, which could be disturbed by extensive damage, sarcopenia, cachexia, or genetic diseases like dystrophy. Many lines of evidence showed that the level of oxygen regulates the course of cell proliferation and differentiation.MethodsIn the present study, we analyzed hypoxia impact on human and pig bone marrow-derived mesenchymal stromal cell (MSC) and mouse myoblast proliferation, differentiation, and fusion. Moreover, the influence of the transplantation of human bone marrow-derived MSCs cultured under hypoxic conditions on skeletal muscle regeneration was studied.ResultsWe showed that bone marrow-derived MSCs increased VEGF expression and improved myogenesis under hypoxic conditions in vitro. Transplantation of hypoxia preconditioned bone marrow-derived MSCs into injured muscles resulted in the improved cell engraftment and formation of new vessels.ConclusionsWe suggested that SDF-1 and VEGF secreted by hypoxia preconditioned bone marrow-derived MSCs played an essential role in cell engraftment and angiogenesis. Importantly, hypoxia preconditioned bone marrow-derived MSCs more efficiently engrafted injured muscles; however, they did not undergo myogenic differentiation.

Highlights

Skeletal muscle regeneration is a complex process that allows restoration of skeletal muscle homeostasis lost due to the injury, such as intensive exercise, surgical procedures, and diseases
The proliferation, migration, and fusion of human and pig bone marrow‐derived MSCs, mouse primary myoblasts, and C2C12 under normoxic and hypoxic conditions First, we analyzed the proliferation of human bone marrow-derived mesenchymal stromal cells, fetal pig bone marrow-derived mesenchymal stromal cells, as well as mouse primary myoblasts, which we used to set up the co-culture experiments (Fig. 1A)
We performed similar analyzes of human bone marrow-derived mesenchymal stromal cells (hMSC) or pig bone marrow-derived mesenchymal stem cells (pMSC) co-cultured with mouse primary myoblasts (mPM) or mouse C2C12 myoblasts

Summary

Introduction

Skeletal muscle regeneration is a complex process that allows restoration of skeletal muscle homeostasis lost due to the injury, such as intensive exercise, surgical procedures, and diseases. The phase of skeletal muscle repair covers myofiber regeneration which is possible due to satellite cells (SCs)—skeletal muscle-specific stem cells, characterized by a PAX7 transcription factor. These cells are tightly connected to the myofibers and located between basal lamina and sarcolemma. The satellite cells remain quiescent and localized between myofiber sarcolemma and basal lamina They are activated in response to muscle injury, proliferate, differentiate into myoblasts, and recreate myofibers. Many lines of evidence showed that the level of oxygen regulates the course of cell prolifera‐ tion and differentiation

Methods

Results

Discussion

Conclusion