In Vitro Cellular & Developmental Biology - Animal ( IF 2.1 ) Pub Date : 2020-10-02 , DOI: 10.1007/s11626-020-00501-1 Vijay Bhaskar Reddy Konala 1, 2 , Ramesh Bhonde 3 , Rajarshi Pal 1
Human mesenchymal stromal cells (MSCs) are currently the leading candidate for cell-based therapeutics. While the use of MSCs in transplantation therapies is widely expanding, still, there is a lot of scope for better understanding of the mechanisms underlying their effects. We have generated MSCs from pre- and post-natal human tissue sources such as Wharton’s jelly (WJ), stem cells from human exfoliated deciduous teeth (SHED), and bone marrow (BM). We then expanded, banked, and characterized them based on morphology, growth kinetics, senescence, immunophenotype, gene expression, and secretion of growth factors. Although the immunophenotype was very similar across MSCs from the three types of donor tissues, they showed minor variations in their growth kinetics. Further, a higher percentage of senescent cells were observed in BM-MSCs than in WJ-MSCs and SHED. Gene expression analysis showed the increased expression of INF-γ, PDGFA, VEGF, IL10, and SDF in SHED over WJ-MSC and BM-MSC. Comparative secretome profiling by ELISA demonstrated the presence of FGF-2, IL-10, PDGF, SDF-1, Ang-1, TGF-β3, HGF, INF-γ, VEGF, and IL-6 in cell culture supernatants. Based on our findings, WJ-MSC and SHED appear more potent than BM-MSC for managing inflammation, immunomodulation, angiogenesis, fibrosis, and scarring. Due to widespread application of MSCs in cell replacement therapy, these subtle differences need to be taken into consideration while designing stem cell–based clinical trials.
中文翻译:
从三种组织来源分离的间充质基质细胞 (MSC) 的分泌组研究揭示了效力的细微差异
人类间充质基质细胞 (MSCs) 目前是基于细胞疗法的主要候选者。尽管 MSC 在移植疗法中的应用正在广泛扩展,但仍然有很多空间可以更好地了解其作用的潜在机制。我们已经从出生前和出生后的人体组织来源,如沃顿氏果冻 (WJ)、人脱落乳牙 (SHED) 和骨髓 (BM) 的干细胞中生成了 MSC。然后,我们根据形态学、生长动力学、衰老、免疫表型、基因表达和生长因子的分泌对它们进行了扩展、存储和表征。尽管来自三种类型供体组织的 MSC 的免疫表型非常相似,但它们的生长动力学差异很小。更远,在 BM-MSC 中观察到的衰老细胞百分比高于 WJ-MSC 和 SHED。基因表达分析表明WJ-MSC 和 BM-MSC 在 SHED 中的INF-γ、PDGFA、VEGF、IL10和SDF。通过 ELISA 进行的比较分泌组分析表明,细胞培养上清液中存在 FGF-2、IL-10、PDGF、SDF-1、Ang-1、TGF-β3、HGF、INF-γ、VEGF 和 IL-6。根据我们的研究结果,WJ-MSC 和 SHED 在控制炎症、免疫调节、血管生成、纤维化和瘢痕形成方面似乎比 BM-MSC 更有效。由于 MSCs 在细胞替代疗法中的广泛应用,在设计基于干细胞的临床试验时需要考虑这些细微的差异。