Deciphering The Transcriptional Activities of Genes Coding For Adipokines and Their Receptors in Porcine Ex Situ-Protected Mesenchymal Stem Cells Undergoing Adipogenic Differentiation

Abstract

Abstract The aim of this study was to investigate the impact of the source of mesenchymal stem cells (MSCs) and their adipogenic derivatives on the relative abundances (RAs) noticed for mRNA transcripts of the selected adipokines (adiponectin and leptin) and their receptors. MSCs were isolated from bone marrow (BM) and subcutaneous adipose tissue (AT ) samples collected post mortem from a total of four gilts (each at the age of 6 months). The stemness of the MSCs was proven via recognizing their abilities to differentiate into adipocytes, osteoblasts, and chondrocytes. By using real-time PCR (RT -qPCR), the quantitative levels of transcriptional activity pinpointed for the adiponectin (ADIPOQ), adiponectin receptor 1 (ADIPOR1), adiponectin receptor 2 (ADIPOR2), leptin (LEP), and leptin receptor (LEPR) genes were comparatively analyzed between non-differentiated AT -MSCs and BM-MSCs and their cell counterparts undergoing differentiation into adipocytes. A significantly higher RA for ADIPOR1 transcripts was identified in the AT-MSCs compared to the BM-MSCs and their adipogenic derivatives (P≤0.05). Moreover, the quantitative levels of LEPR transcripts were shown to increase significantly among adipocytes originating from differentiated BM-MSCs compared to undifferentiated BM-MSCs, AT -MSCs, and AT -MSC-derived adipocytes (P≤0.05). Nonetheless, there was no significant inter-group variability in the RAs of the ADIPOQ, ADIPOR2, or LEP mRNA transcripts (P>0.05). Cumulatively, the in vitro models focused on the identification and detailed exploration of transcriptomic signatures of undifferentiated pig BM- and AT -MSCs, and the molecular mechanisms underlying the adipogenic differentiation pathways of the above-indicated two distinct sources of stem cells were developed and optimized in the current investigation for the very first time. These ex vivo porcine models might confirm the expedited functional mobilization of ex situ-protected MSCs and their enhanced capacity to be transcriptionally reprogrammed into adipocytes due to physiopathological alterations in the expression profiles of adipokines and their receptors, which are prompted and progressed in obese and superobese women patients at the peri-pubertal period of ontogenesis. Finally, the models providing comprehensive molecular insights into ex situ-protected porcine BM- and AT -derived MSCs and their differentiated derivatives can be largely suitable for the biotechnologically assisted rescue and restitution of endangered representatives of rare native breeds of domestic pig.