Retinoic acid induces the osteogenic differentiation of cat adipose tissue‐derived stromal cells from distinct anatomical sites

Abstract

Mesenchymal stromal cells-based regenerative orthopedic therapies have been usedin cats as a promising and innovative therapeutic approach to enhance the repair of bone defects. Adipose tissue-derived stromal cells (ADSCs) can be obtained from two main sites-subcutaneous and visceral-with established differences regarding structure, composition, cell content, and functionality.

The Osteogenic Potential of Falciform Ligament-Derived Stromal Cells—A Comparative Analysis between Two Osteogenic Induction Programs

“…In a comparative approach relative to cultures grown in the presence of the osteogenic inducers, more elongated cell morphology and an increased cytoplasmic area were verified in cells cultured in RA, compared to those cultured in DEX. A similar trend was observed previously in studies with cat- and mouse-derived cells [ 30 , 66 ]. Despite the fact that the mechanism by which cells cultured in RA exhibit the reported differences is not fully described, it is thought to be related to alterations in the cytoskeletal arrangement, attributed to an increased thickness of actin stress fibers promoted by the MAPK signaling activation [ 67 , 68 ].…”

“…Contrariwise, in the presence of RA, a reduction in the proliferation is expected, which is thought to be related to a decreased expression in Cdk2, Cdk4/Cdk6, D-type cyclins (D1, D2, and D3), and cyclin E, which suppress the progression from the G1 to the S phase, thus hindering cell proliferation [ 56 , 59 ]. A similar trend has been reported in studies conducted on ADSCs from distinct species exposed to RA, namely those grown from rats, cows, and cats [ 30 , 31 , 57 ].…”

“…ADSCs’ cultures, regardless of the tissue of origin, exhibited a fibroblastic-like arrangement with an elongated morphology, and well-marked F-actin organized into evident stress fibers when grown in basal conditions. This morphology is characteristic of ADSCs obtained from dogs [ 2 , 3 , 58 , 60 ] and other species, namely rats, mice, cats, horses, rabbits, pigs, and humans [ 3 , 30 , 61 , 62 , 63 , 64 , 65 ]. In a comparative approach relative to cultures grown in the presence of the osteogenic inducers, more elongated cell morphology and an increased cytoplasmic area were verified in cells cultured in RA, compared to those cultured in DEX.…”

“…Adipose tissue samples were decontaminated, as previously described [ 30 ]. Subsequently, samples were cut into 1–3 mm 3 fragments and digested for 40 min, at 37 °C, with 0.1% collagenase type IA (Sigma-Aldrich) in phosphatase-buffered saline (PBS; Sigma-Aldrich).…”

“…Since then, most applications, either experimental or within the frame of therapeutic applications, have relied on osteogenic induction through the supplementation of the culture medium with dexamethasone, β-glycerophosphate, and ascorbic acid [ 2 , 13 , 29 ]. Most recently, several studies with rat-, mouse-, and cat-derived cells have reported the ability of retinoic acid (RA) to promote osteogenic differentiation [ 30 , 31 , 32 ]. In dog-derived cells, so far, only one study evaluated the RA-mediated osteogenic induction of ADSCs, focusing exclusively on cells from the subcutaneous depot [ 33 ], with no available data on visceral-derived populations.…”