Heimann Manuela, Elashry Mohamed I, Klymiuk Michele C, Eldaey Asmaa, Wenisch Sabine, Arnhold Stefan
Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, Germany.
Clinic of Small Animals, c/o Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, Germany.
Animals (Basel). 2023 Oct 15;13(20):3224. doi: 10.3390/ani13203224.
The investigation of adipose tissue-derived mesenchymal stem cells (ASCs) has received considerable interest in regenerative medicine. A nontoxic adipogenic induction protocol valid for cells of different mammalian species has not been described. This study aims to establish an adipogenic differentiation protocol suitable for horses, sheep, dogs, murines, and human cells. An optimized rosiglitazone protocol, consisting of 5% fetal calf serum in Dulbecco's Modified Eagle's Medium, 10 μg/mL insulin, 0.55 μg/mL transferrin, 6.8 ng sodium selenite, 1 μM dexamethasone, and 1-5 μM of rosiglitazone, is compared to the 3-isobutyl-1-methylxantine (IBMX) protocol, where rosiglitazone was replaced with 0.5 mM IBMX and 0.2 mM indomethacin. Cell viability, cytotoxicity, a morphometric analysis of the lipid, and the expression of adipogenic markers for 14 days were assessed. The data revealed that using 5 µM of rosiglitazone promotes the adipogenic differentiation capacity in horse, sheep, and dog cells compared to IBMX induction. Meanwhile, marked reductions in the cell viability and cell number with the IBMX protocol were detected, and rosiglitazone increased the cell number and lipid droplet size, prevented apoptosis, and upregulated and expression in the cells of most of the species. Our data revealed that the rosiglitazone protocol improves the adipogenesis of ASCs, together with having less toxicity, and should be considered for cell reproducibility and clinical applications targeting obesity.
脂肪组织来源的间充质干细胞（ASCs）的研究在再生医学领域引起了广泛关注。目前尚未有适用于不同哺乳动物物种细胞的无毒成脂诱导方案被报道。本研究旨在建立一种适用于马、羊、狗、小鼠和人类细胞的成脂分化方案。将优化后的罗格列酮方案（即在杜氏改良 Eagle 培养基中添加 5%胎牛血清、10 μg/mL 胰岛素、0.55 μg/mL 转铁蛋白、6.8 ng 亚硒酸钠、1 μM 地塞米松和 1 - 5 μM 罗格列酮）与 3 - 异丁基 - 1 - 甲基黄嘌呤（IBMX）方案进行比较，在 IBMX 方案中，罗格列酮被 0.5 mM IBMX 和 0.2 mM 吲哚美辛取代。评估了细胞活力、细胞毒性、脂质的形态计量分析以及 14 天内成脂标志物的表达。数据显示，与 IBMX 诱导相比，使用 5 μM 罗格列酮可促进马、羊和狗细胞的成脂分化能力。同时，检测到 IBMX 方案会导致细胞活力和细胞数量显著降低，而罗格列酮增加了细胞数量和脂滴大小，防止细胞凋亡，并上调了大多数物种细胞中的 和 表达。我们的数据表明，罗格列酮方案可改善 ASCs 的成脂作用，且毒性较小，在细胞可重复性和针对肥胖症的临床应用方面应予以考虑。
