article · International Journal of Molecular Sciences
Mesenchymal stem cells serve as valuable candidates for regenerative medicine and cellular therapies. This research established isolation and culture procedures for mesenchymal stem cells sourced from five distinct canine tissues: adipose tissue, bone marrow, placenta, umbilical cord, and amniotic membrane. All five cell types exhibited characteristic fibroblast-like growth, tested positive for the surface marker CD44, and lacked CD34 expression. Each variety successfully differentiated into fat and bone cells in vitro, with third-generation cultures demonstrating the highest rates of proliferation. Adipose-derived cells displayed the fastest population doubling time of 15.8 hours and differentiated into bone and fat lineages more rapidly than the other types. Comparative transcriptome profiling revealed that adipose and bone marrow cells shared the highest degree of genetic homology, whereas placenta-derived cells showed distinct expression patterns.
Identifying optimal sources for stem cells is essential for advancing veterinary regenerative medicine. By profiling how canine stem cells from different tissues proliferate and differentiate, this work helps clinicians and researchers select the most effective cell types for therapeutic development. The findings highlight adipose tissue as an exceptionally fast-growing and versatile candidate for future animal cell therapies.
This research provides baseline comparative data to inform cell selection for canine cellular therapies and tissue engineering applications. Potential users include veterinary biopharmaceutical developers and regenerative medicine clinics. Because the work remains at the stage of in vitro characterisation and transcriptomic profiling, it represents early-stage laboratory research that requires extensive in vivo safety and efficacy testing before reaching commercial clinical use.
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Mesenchymal stem cells (MSCs) are the most promising seed cells for cell therapy. Comparing the biological and transcriptome gene characteristics of MSCs from different sources provides an important basis for the screening of clinically used cells. The main purpose of this experiment was to establish methods for the isolation and culture of MSCs from five different canine sources, including adipose tissue, bone marrow, umbilical cord, amniotic membrane, and placenta, and compare biological and transcriptome characteristics of MSCs, in order to provide a basis for the clinical application of canine MSCs. MSCs were isolated from Chinese pastoral dogs, and the following experiments were performed: (1) the third, sixth, and ninth generations of cells were counted, respectively, and a growth curve was plotted to calculate the MSC population doubling time; (2) the expression of CD34 and CD44 surface markers was studied by immunofluorescence; (3) the third generation of cells were used for osteogenetic and adipogenic differentiation experiments; and (4) MSC transcriptome profiles were performed using RNA sequencing. All of the five types of MSCs showed fibroblast-like adherent growth. The cell surface expressed CD44 instead of CD34; the third-generation MSCs had the highest proliferative activity. The average population doubling time of adipose mesenchymal stem cells (AD-MSCs), placenta mesenchymal stem cells (P-MSCs), bone marrow mesenchymal stem cells (BM-MSCs), umbilical cord mesenchymal stem cells (UC-MSCs), and amniotic mesenchymal stem cells (AM-MSCs) were 15.8 h, 21.2 h, 26.2 h, 35 h, and 41.9 h, respectively. All five types of MSCs could be induced to differentiate into adipocytes and osteoblasts in vitro, with lipid droplets appearing after 8 days and bone formation occurring 5 days after AD-MSC induction. However, the multilineage differentiation for the remaining of MSCs was longer compared to that of the AD-MSCs. The MSC transcriptome profiles showed that AD-MSC and BM-MSCs had the highest homology, while P-MSCs were significantly different compared to the other four types of MSCs. All the isolated MSCs had the main biological characteristics of MSCs. AD-MSCs had the shortest time for proliferation, adipogenesis, and osteogenic differentiation.
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DOI: 10.3390/ijms20061485
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