研究了血小板裂解物(PL)作为细胞培养中胎牛血清(FBS)的潜在替代品。然而,关于其对马间充质基质细胞(eMSCs)免疫谱的影响的研究有限。本研究旨在评估不同PL制剂对增殖能力的影响,多潜力,和马脂肪组织来源的MSCs(eAD-MSCs)的免疫谱。在三种培养条件下评估了eAD-MSCs(n=7)的体外生长动力学和三系分化:培养基浓度PL(MPL),高浓度PL(HPL),和FBS作为控制。通过研究免疫原性受体如MHCI,MHCII,和免疫调节分子IL-6,IL-10和TNF-α,由基因表达决定,表面标记表达,和细胞因子定量。两种PL配方,来自5个捐赠者,MPL和HPL的血小板计数比基线血浆高3.3和6.5倍,分别。与基线相比,在两种PL制剂中发现更高浓度的TGF-β和PDGF。此外,MPL和HPL传代培养显示增殖,克隆,和类似于FBS的多能容量。PL培养细胞的免疫谱表现出与参考条件相似的与免疫原性和免疫调节相关的基因表达水平。MHCII的表面抗原存在也相似。然而,HPL培养基在培养上清液中显示出较高的IL-6、IL-10和TNF-α浓度。总之,与FBS相比,两种PL培养基都含有更高浓度的生长因子,支持eAD-MSCs增殖的体外培养,克隆,和类似于参考培养基的多能容量。尽管如此,PL的使用导致免疫调节细胞因子微环境的变化,与MPL和FBS相比,HPL培养基中IL-6、IL-10和TNF-α的浓度更高。
Platelet lysate (PL) is investigated as a potential replacement for fetal bovine serum (FBS) in cell culture. However, there is limited research on its impact on the immune profile of equine mesenchymal stromal cells (eMSCs). This study aimed to evaluate the effects of different PL formulations on the proliferative capacity, multipotentiality, and immune profile of equine adipose tissue-derived MSCs (eAD-MSCs). In vitro growth kinetics and trilineage differentiation of eAD-MSCs (n = 7) were assessed under three culture conditions: medium-concentration PL (MPL), high-concentration PL (HPL), and FBS as a control. The immune profile was evaluated by studying the expression of immunogenic receptors such as MHC I, MHC II, and immunomodulatory molecules IL-6, IL-10, and TNF-α, determined by gene expression, surface marker expression, and cytokine quantification. Both PL formulations, pooled from 5 donors, exhibited 3.3 and 6.5-fold higher platelet counts than baseline plasma for MPL and HPL, respectively. Higher concentrations of TGF-β and PDGF were found in both PL formulations compared to baseline. Furthermore, MPL and HPL subcultures demonstrated proliferative, clonogenic, and multipotent capacities similar to FBS. The immune profile of PL-cultured cells exhibited gene expression levels related to immunogenicity and immunomodulation similar to the reference condition, and the surface antigen presence of MHC II was also similar. However, HPL media exhibited higher IL-6, IL-10, and TNF-α concentrations in the culture supernatant. In conclusion, both PL media contained higher concentrations of growth factors compared to FBS, supporting the in vitro culture of eAD-MSCs with proliferative, clonogenic, and multipotent capacity similar to the reference medium. Nonetheless, PL usage led to a variation in the immunomodulatory cytokine microenvironment, with higher concentrations of IL-6, IL-10, and TNF-α in HPL media compared to MPL and FBS.