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Transplantation of Induced Pluripotent Stem Cell-derived Mesoangioblast-like Myogenic Progenitors in Mouse Models of Muscle Regeneration
Published on: January 20, 2014
Resetting the problem of cell death following muscle-derived cell transplantation: detection, dynamics and mechanisms
Daniel Skuk1, Nicolas J Caron, Marlyne Goulet
1Unité de recherche en Génétique humaine, Centre de Recherche du Centre Hospitalier de l'Université Laval, CHUL du CHUQ, Ste-Foy, Québec, Canada. Daniel.Skuk@anm.ulaval.ca
Abstract:
We conducted a study in mice to reevaluate and clarify many aspects of the early survival of muscle cells following transplantation. Male mouse muscle cells (primary-cultures and T-antigen-immortalized clones) labeled with [14C]thymidine and beta-galactosidase were injected into female muscles. Each label was detected in the muscles after different time periods. TUNEL, alizarin red, and immunodetection of active caspase-3 were done in muscle sections. The donor cell labels disappeared from the muscles following donor cell death, but this was not instantaneous and even if the donor cells were killed before transplantation, the first 6 hours were not enough to clear [14C]thymidine and Y chromosome. Using the cell pellet before injection as the 100% baseline for cells injected to evaluate cell death can lead to misinterpretations: the Y-chromosome band was 5-fold stronger than that of a muscle injected with cells, irrespective of whether the cells were previously killed or not. There was no evidence of an immediate massive donor cell death. Necrosis (detected by alizarin red) and apoptosis (detected by active caspase-3) were present among the donor myoblasts following transplantation. Necrosis seemed to be the most important mechanism during the first hours. T-antigen immortalized cells died earlier and more massively than primary-cultured cells, but the surviving cells proliferated more. Indeed, they seemed to exhibit more apoptosis and they triggered a more rapid CD8+ cell infiltration. As a result of our findings, many concepts concerning the early donor cell death following myoblast transplantation must be reconsidered.
Insights
Early muscle cell survival after transplantation is complex. Donor cell death, via necrosis and apoptosis, is not immediate and requires reevaluation of current concepts in myoblast transplantation.
Area of Science:
- Muscle cell biology
- Regenerative medicine
- Transplantation immunology
Background:
- Understanding early donor cell survival is crucial for optimizing myoblast transplantation therapies.
- Previous studies have suggested rapid and massive donor cell death post-transplantation, but this requires further investigation.
Purpose of the Study:
- To reevaluate and clarify early muscle cell survival following transplantation in mice.
- To investigate the mechanisms and kinetics of donor cell death after myoblast transplantation.
Main Methods:
- Male mouse muscle cells (primary cultures and T-antigen immortalized clones) were labeled and injected into female recipient muscles.
- Detection of donor cell labels ([14C]thymidine, beta-galactosidase, Y chromosome) over time.
- Assessment of cell death using TUNEL assay, alizarin red staining (necrosis), and active caspase-3 immunodetection (apoptosis).
Main Results:
- Donor cell labels did not disappear instantaneously after cell death, with [14C]thymidine and Y chromosome persisting for hours even in pre-killed cells.
- Alizarin red staining indicated necrosis as a primary mechanism of cell death in the early hours post-transplantation.
- T-antigen immortalized cells exhibited more rapid and massive death but also showed increased proliferation and triggered faster CD8+ T-cell infiltration compared to primary cells.
Conclusions:
- Established concepts regarding immediate massive donor cell death following myoblast transplantation need reconsideration.
- Both necrosis and apoptosis contribute to donor cell loss, with necrosis being dominant initially.
- Immortalized cells present a different survival and death profile compared to primary cells, impacting transplantation outcomes.
