Immunophenotypic expression by flow cytometric analysis of cocultured skeletal muscle and bone marrow mesenchymal stem cells for therapy into myocardium.
The product generated by skeletal muscle and bone marrow mesenchymal stem cell cocultures has been demonstrated to improve the functional outcomes after cell therapy in postinfarction or Chagas myocardiopathy. This coculture method allows cell interactions in vitro, diminishing the operational costs of the culture/expansion as well as leading to angiogenesis and myogenesis for regeneration of the injured heart. Flow cytometric analysis may better characterize the cellular types in this model. Our objective was to use flow cytometry to analyze the immunophenotype expressed in this coculture model. The coculture was performed in accordance with Carvalho for 21 days. Flow cytometry was performed before and after coculture to characterize the immunophenotypic profile of cellular subsets, namely, the surface markers CD31, CD34, CD44H, CD45, CD49d, CD54, CD73, CD90, CD105, CD106, Myo-D, M-cadherin, and Connexin-43. Statistics were performed by the nonparametric Friedman test (P < .05) with post-hoc analysis by the nonparametric Wilcoxon test (P < or = .017, Bonferroni correction). The results demonstrated statistical significance for CD45(+) in 89.49% of mononuclear cells, 3.58% in skeletal muscle cells, and 4.74% among cocultured cells (P = .0094); and CD90(+) in 36.18% of mononuclear cells, 6.01% in skeletal muscle cells, and 48.94% among cocultured cells (P = .0420). The cocultured cells expressed the markers CD73(++), CD90(+++), CD45(-), CD34(+), CD105(-/+), CD106(-/+), M-cadherin(-/+), and Connexin-43(-/+). In conclusion, flow cytometric analysis showed a heterogeneous adherent cell population in this coculture model.
Carvalho
K A T C
KA
Cellular Culture Experimental Laboratory, PUCPR, Curitiba, Brazil. katherinecarv@gmail.com
Oliveira
L
L
Malvezzi
M
M
Simeoni
R B
RB
Francisco
J C
JC
Olandoski
M
M
Guarita-Souza
L C
LC
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D018920:Coculture Techniques; D005434:Flow Cytometry / Q000379:methods; D006321:Heart / Q000502:physiology; D006331:Heart Diseases / Q000601:surgery*; D016130:Immunophenotyping / Q000379:methods*; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D018482:Muscle, Skeletal / Q000166:cytology* / Q000502:physiology; D009206:Myocardium / Q000166:cytology; D051381:Rats