Cardiac Bmi1(+) cells contribute to myocardial renewal in the murine adult heart.
BACKGROUND
The mammalian adult heart maintains a continuous, low cardiomyocyte turnover rate throughout life. Although many cardiac stem cell populations have been studied, the natural source for homeostatic repair has not yet been defined. The Polycomb protein BMI1 is the most representative marker of mouse adult stem cell systems. We have evaluated the relevance and role of cardiac Bmi1 (+) cells in cardiac physiological homeostasis.


METHODS
Bmi1 (CreER/+);Rosa26 (YFP/+) (Bmi1-YFP) mice were used for lineage tracing strategy. After tamoxifen (TM) induction, yellow fluorescent protein (YFP) is expressed under the control of Rosa26 regulatory sequences in Bmi1 (+) cells. These cells and their progeny were tracked by FACS, immunofluorescence and RT-qPCR techniques from 5 days to 1 year.


RESULTS
FACS analysis of non-cardiomyocyte compartment from TM-induced Bmi1-YFP mice showed a Bmi1 (+)-expressing cardiac progenitor cell (Bmi1-CPC: B-CPC) population, SCA-1 antigen-positive (95.9 ± 0.4 %) that expresses some stemness-associated genes. B-CPC were also able to differentiate in vitro to the three main cardiac lineages. Pulse-chase analysis showed that B-CPC remained quite stable for extended periods (up to 1 year), which suggests that this Bmi1 (+) population contains cardiac progenitors with substantial self-maintenance potential. Specific immunostaining of Bmi1-YFP hearts serial sections 5 days post-TM induction indicated broad distribution of B-CPC, which were detected in variably sized clusters, although no YFP(+) cardiomyocytes (CM) were detected at this time. Between 2 to 12 months after TM induction, YFP(+) CM were clearly identified (3 ± 0.6 % to 6.7 ± 1.3 %) by immunohistochemistry of serial sections and by flow cytometry of total freshly isolated CM. B-CPC also contributed to endothelial and smooth muscle (SM) lineages in vivo.


CONCLUSIONS
High Bmi1 expression identifies a non-cardiomyocyte resident cardiac population (B-CPC) that contributes to the main lineages of the heart in vitro and in vivo.
Cardiovascular Development and Repair Department, Spanish National Cardiovascular Research Center (CNIC), Madrid, Spain. inigo.valiente.alandi@cchmc.org.|The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA. inigo.valiente.alandi@cchmc.org.
Iñigo
I
Valiente-Alandi
Cardiovascular Development and Repair Department, Spanish National Cardiovascular Research Center (CNIC), Madrid, Spain. Calbo@vivebiotech.com.|Vivebiotech, San Sebastian, Spain. Calbo@vivebiotech.com.
Carmen
C
Albo-Castellanos
Cardiovascular Development and Repair Department, Spanish National Cardiovascular Research Center (CNIC), Madrid, Spain. dherrero@cnb.csic.es.|Immunology and Oncology Department, Spanish National Center for Biotechnology (CNB-CSIC), Madrid, Spain. dherrero@cnb.csic.es.
Diego
D
Herrero
Microscopy Unit, Spanish National Cardiovascular Research Center (CNIC), Madrid, Spain. earza@cnic.es.
Elvira
E
Arza
Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT)- Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain. m.garcia.gomez@ciemat.es.|Advanced Therapies Mixed Unit, Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD, UAM), Madrid, Spain. m.garcia.gomez@ciemat.es.
Maria
M
Garcia-Gomez
Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT)- Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain. jc.segovia@ciemat.es.|Advanced Therapies Mixed Unit, Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD, UAM), Madrid, Spain. jc.segovia@ciemat.es.
José C
JC
Segovia
Howard Hughes Medical Institute University of Utah, Salt Lake City, UT, USA. mario.capecchi@genetics.utah.edu.
Mario
M
Capecchi
Cardiovascular Development and Repair Department, Spanish National Cardiovascular Research Center (CNIC), Madrid, Spain. abernad@cnb.csic.es.|Immunology and Oncology Department, Spanish National Center for Biotechnology (CNB-CSIC), Madrid, Spain. abernad@cnb.csic.es.
Antonio
A
Bernad
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D006321:Heart / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D000066450:Mouse Embryonic Stem Cells / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D063150:Polycomb Repressive Complex 1 / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D017208:Rats, Wistar; D012038:Regeneration
D015415:Biomarkers; C489421:Bmi1 protein, mouse; D011518:Proto-Oncogene Proteins; D063150:Polycomb Repressive Complex 1