Induced pluripotent stem cell-derived cardiomyocytes proliferate by exposure to conditioned medium from mesenchymal stem cells

NATALIA LUCÍA SANTÍN VELAZQUE; MARIA AGUSTINA SCARAFIA; ALAN MIQUEAS MÖBBS; MARIA CELESTE BIANI; ANTONELLA LOMBARDI; CYNTHIA ABAN; CAROLINA COLLI; ARIEL WAISMAN; LUCIA NATALIA MORO; ALEJANDRO DAMIÁN LA GRECA; GUSTAVO EMILIO SEVLEVER; CARLOS DANIEL LUZZANI; SANTIAGO GABRIEL MIRIUKA

Mar del Plata

Congreso; LXIII Annual Meeting of the Argentine Society for Clinical Investigation (SAIC); 2018

SAIC - Sociedad Argentina de Investigacion Clinica

Inhuman heart, shortly after birth, cardiomyocytes (CM) undergo onelast round of division followed by escape from cell cycle. Productionof induced pluripotent stem cell-derived CM (iPSC-CM) is apotentially promising strategy for regenerative therapies.Cardiomyocytes in adult mammalian heart are characterized by lack ofproliferation, thus shaping a growing interest in identifying factorsrelevant in the regulation of iPSC-CMs cell cycle. In recent years,accumulating evidences highlighted the regenerative properties ofmesenchymal stem cells (MSC), a well-known type of multipotent cell.Particularly, their secretome has considerable pro-mitotic potential.Hence, the aim of this work was to study iPSC-CM ability to re-entercell cycle after exposure to conditioned medium from MSC. iPSC-CMswere obtained by the implementation of a previously reported protocoland cell proliferation was monitored by stable introduction of afluorescent ubiquitination cell cycle indicator (FUCCI) in iPSC(Fucci-iPSC). FUCCI construction consists of mCherry and mVenusfluorescent proteins fused to different regulators of cell cycle:cdt1 and geminin, respectively. Shift in color from red to greenallow us to follow and quantify cell division events. Fucci-iPSC-CMswere treated or not with 48h-conditioned medium for 24, 48 and 72h.Images were captured in fluorescent microscope. Number of greenfluorescent cells increased from 15%±2.36 to 29%±12.76 in the first24h and from 5%±2.94 to 23%±4.23 at 48h. Differential proliferationrates were only significant at 72h of treatment with conditionedmedium, showing a change from 5%±2.8 to 23%±2.07 (p