Virology Department, Croatian Veterinary Institute, Savska Cesta 143, 10 000 Zagreb, Croatia.
Department of Veterinary Pathology, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10 000 Zagreb, Croatia.
Int J Mol Sci. 2021 Jul 12;22(14):7476. doi: 10.3390/ijms22147476.
The influence of cultivation on the expression pattern of canine adipose-derived mesenchymal stem cells (cAD-MSCs) surface markers, contributing to, among others, the promotion of growth, proliferation, differentiation and immunomodulatory mechanisms of an excellent therapeutic, is still unknown. To fill the gap, we investigated CD90, CD44, CD73, CD29, CD271, CD105, CD45 and CD14 patterns of expression at the protein level with flow cytometry and mRNA level using a real-time polymerase chain reaction array. Gentle variations of expression occurred during cultivation, along with increased CD90, CD44 and CD29 expression, low and decreasing CD271 and CD73 expression and a decrease of initially high CD105. As expected, CD45 and CD14 were not expressed by cAD-MSCs. Interestingly, we discovered a significant decrease of CD73 expression, compared to early (P1-P3) to late (P4-P6) passages, although the CD73 gene expression was found to be stable. The percentage of positive cells was found to be higher for all positive markers up to P4. As CD73's one important feature is a modulation from a pro-inflammatory environment to an anti-inflammatory milieu, the expression of CD73 in our conditions indicate the need to consider the time cells spend in vitro before being transplanted into patients, since it could impact their favourable therapeutical properties.
培养对犬脂肪间充质干细胞(cAD-MSCs)表面标志物表达模式的影响,有助于促进其生长、增殖、分化和免疫调节机制,这一点尚不清楚。为了填补这一空白,我们使用流式细胞术检测了 CD90、CD44、CD73、CD29、CD271、CD105、CD45 和 CD14 的蛋白水平表达模式,并使用实时聚合酶链反应阵列检测了 mRNA 水平表达模式。在培养过程中,表达水平发生了轻微变化,同时 CD90、CD44 和 CD29 的表达增加,CD271 和 CD73 的表达降低,CD105 的表达最初较高,但逐渐降低。不出所料,cAD-MSCs 不表达 CD45 和 CD14。有趣的是,与早期(P1-P3)相比,我们发现 CD73 的表达在晚期(P4-P6)明显降低,尽管 CD73 基因的表达是稳定的。在 P4 之前,所有阳性标志物的阳性细胞百分比都较高。由于 CD73 的一个重要特征是从促炎环境向抗炎环境的调节,因此在我们的条件下 CD73 的表达表明需要考虑细胞在被移植到患者体内之前在体外培养的时间,因为这可能会影响它们有利的治疗特性。