Timing of induction of cardiomyocyte differentiation for in vitro cultured mesenchymal stem cells: a perspective for emergenciesThis article is one of a selection of papers from the NATO Advanced Research Workshop on Translational Knowledge for Heart Health (published in part 1 of a 2-part Special Issue).

Abstract

Mesenchymal stem cells (MSCs) have the capacity to differentiate into osteoblasts, chondrocytes, adipocytes, myocytes, and cardiomyocytes. Several established methods are presently available for in vitro isolation of MSCs from bone marrow. However, the duration necessary to culture them can be a major handicap to cell-based therapies needed for such urgent cardiovascular conditions as acute myocardial infarction and acute hindlimb ischemia. The best timing of cardiomyocyte differentiation induction after MCS isolation and expansion is still an unresolved issue. Our goal was to investigate the possibility of obtaining functional cardiomyocytes from rat MSC within a shorter time period. We examined MSCs' colony-forming capacity, CD90 and CD34 immunoreactivity during the 14 days of culturing. Cardiomyocyte differentiation was induced by 5-azacytidine. Immunohistochemic staining, together with intracellular Ca 2+ measurement experiments, revealed that MSCs do not differentiate into any specific cell lineage but show the characteristics of MSCs on both the 9th and 14th days of the culture. To check the potential for differentiation into cardiomyocytes, experiments with caffeine application and depolarization with KCl were performed. The cells possessed some of the specific biochemical features of contracting cells, with slightly higher capacities on the 14th day. Cells from 9th and 14th days of the culture that were treated with 5-azacytidine had a higher expression of cardiac-specific markers such as troponin I, a-sarcomeric actin, and MEF2D compared with the control groups. This study illustrates that it is possible to get functional cardiomyocytes from in vitro MSC culture in a shorter time period than previously achieved. This reduction in time may provide emergency cases with access to cell-based therapies that may have previously been unavailable.Key words: mesenchymal stem cells, cardiomyocytes, differentiation, rat, in vitro, gene expression. Résumé :Les cellules souches mésenchymateuses (CSM) ont la capacité de se différencier en ostéoblastes, chondrocytes, adipocytes, myocytes et cardiomyocytes. Plusieurs méthodes éprouvées permettent d'isoler in vitro les CSM de la moelle osseuse. Toutefois, la durée requise pour leur culture pourrait être un désavantage majeur pour la thérapie cellulaire d'urgences cardiovasculaires comme l'infarctus du myocarde et l'ischémie des membres inférieurs. Le meilleur moment de l'induction de la différenciation des cardiomyocytes après l'isolement et l'expansion des CSM n'a toujours pas été déter-miné. Nous avons étudié la possibilité d'obtenir des cardiomyocytes fonctionnels de CSM de rats sur une courte période de temps. Nous avons examiné la capacité de formation de colonies des CSM et l'immunoréactivité à CD90 et CD34 durant les 14 jours de culture. La différenciation en cardiomyocytes a été induite par la 5-azacytidine. La coloration immunohistochimique ainsi que des mesures du Ca 2+ intracellulaire ont révélé que les CSM ne se différencient pas en une lignée cell...

Bone Marrow-Derived Mesenchymal Stem Cells Promote Hepatic Regeneration after Partial Hepatectomy in Rats

“…MSCs, the major stem cells in bone marrow, constitute the microenvironment of the bone marrow, regulate hematopoietic function and differentiate into various cells, including hepatocytes [10,11,22,23,24]. BM-MSCs are easily isolated from bone marrow, readily cultured in vitro and can be used for autologous transplantation.…”

Stem Cell Therapy for Myocardial Infarction: Are We Missing Time?

“…Simply stated, stem cell researchers might focus on other types of stem cells or stem cell origins (autologous vs. heterologous) if the time between MI and stem cell treatment is limited. Researchers are already investigating ways to reduce the culture time of mesenchymal stem cells in emergency situations such as myocardial infarction [61]. …”

Poly (hydroxyethyl methacrylate-glycidyl methacrylate) films modified with different functional groups: In vitro interactions with platelets and rat stem cells

“…Information on the effect of surface chemical composition, especially surface charge, Lewis acid/base donors, and hydrophilic/hydrophobic balance, on MSC adhesion and differentiation is very limited. MSCs not only have potential to differentiate into tissue-specific cell types [13][14][15], but also lack immunogenicity and ethical problems [16]. Thus, MSCs emerge as an important player of cellular-based therapies and new approaches are required to increase the viability and functionality of MSCs.…”