Infoscience

Student project

Isolation and characterisation of a porcine cardiac adherent cell population from heart biopsies

Cardiac diseases represent the major cause of mortality in the Western world. The discovery of the promising human Cardiac Adherent Proliferating (CAP) cells for a potential cardiac cell therapy by the Tissue Engineering group induced the need of a reliable animal model. The aim of the project is to identify a porcine cardiac cell population having similar characteristics as human CAP cells. Here a protocol for isolating cells from porcine heart biopsies have been established, as well as the in vitro expansion method. Additionally, the characterization of those expanded porcine cardiac cells was accomplished by growth kinetics study, FACS analysis, multilineage potential and myogenic induction assays. In this study we identified a not yet described porcine cardiac adherent cell population directly isolated from biopsies within the inner right ventricle. With almost $5\times {10}^{7}$ expanded cells after 30 days in culture, porcine cardiac cells present a high rate of proliferation in vitro. Porcine cardiac cells maintain a swirl-like pattern until 5-6 days in culture, and become fibroblast-like with a parallel organization when confluent. FACS analysis indicated that porcine cells are mainly CD90+ in opposite to human CAP cells. Both species show a similar CD45- and CD44+ profile. The multilineage assay showed no histological differentiation into fat, bone and cartilage on porcine cardiac cells like on human CAP cells. However gene expression analysis on porcine cardiac cells indicated that aP2 was upregulated in population undergoing an adipogenic induction, as well as osteopontine expression in osteogenic induction population. Porcine cardiac cell myogenic induction potential was tested on C2C12 myogenic mouse cells. The mouse cells differentiated in muscle in vitro thanks to the influence of those cardiac cells. Both cell populations have similar characteristics and the main distinction is the different CD90 expression profile. Those results encourage to realise additional studies, to confirm that those isolated cells could be a reliable model of human CAP cells.

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