Wang Dong-Hui, Wu Xue-Mei, Chen Jia-Song, Cai Zhi-Gang, An Jun-Hui, Zhang Ming-Yue, Li Yuan, Li Fei-Ping, Hou Rong, Liu Yu-Liang
Chengdu Research Base of Giant Panda Breeding, 1375 Panda Road, Northern Suburb, Chengdu, 610081, Sichuan Province, China.
Sichuan Key Laboratory of Conservation Biology for Endangered Wildlife, 1375 Panda Road, Northern Suburb, Chengdu, 610081, Sichuan Province, China.
Conserv Physiol. 2022 Feb 21;10(1):coac004. doi: 10.1093/conphys/coac004. eCollection 2022 Jan 1.
Endometrial mesenchymal stem cells (eMSCs) are undifferentiated endometrial cells with self-renewal, multidirectional differentiation and high proliferation potential. Nowadays, eMSCs have been found in a few species, but it has never been reported in endangered wild animals, especially the red panda. In this study, we successfully isolated and characterized the eMSCs derived from red panda. Red panda eMSCs were fibroblast-like, had a strong proliferative potential and a stable chromosome number. Pluripotency genes including , and were highly expressed in eMSCs. Besides, cultured eMSCs were positive for MSC markers CD44, CD49f and CD105 and negative for endothelial cell marker CD31 and haematopoietic cell marker CD34. Moreover, no reference RNA-seq was used to analyse the eMSCs transcriptional expression profile and key pathways. Compared with skin fibroblast cell group, 9104 differentially expressed genes (DEGs) were identified, among which are 5034 genes upregulated, 4070 genes downregulated and the top 20 enrichment pathways of DEGs in Gene Ontology (GO) and the Kyoto Encyclopedia of Genes Genomes (KEGG) mainly associated with G-protein coupled receptor signalling pathway, carbohydrate derivative binding, nucleoside binding, ribosome biogenesis, cell cycle, DNA replication, Ras signalling pathway and purine metabolism. Among the DEGs, some representative genes about promoting MSCs differentiation and proliferation were upregulated and promoting fibroblasts proliferation were downregulated in eMSCs group. Red panda eMSCs also had multiple differentiation ability and could differentiate into adipocytes, chondrocytes and hepatocytes. In conclusion, we, for the first time, isolated and characterized the red panda eMSCs with ability of multiplication and multilineage differentiation . The new multipotential stem cell could be beneficial not only for the germ plasm resources conservation of red panda, but also for basic or pre-clinical studies in the future.
子宫内膜间充质干细胞（eMSCs）是具有自我更新、多向分化和高增殖潜能的未分化子宫内膜细胞。如今，已在少数物种中发现了eMSCs，但在濒危野生动物中从未有过报道，尤其是小熊猫。在本研究中，我们成功分离并鉴定了源自小熊猫的eMSCs。小熊猫eMSCs呈成纤维细胞样，具有很强的增殖潜能且染色体数目稳定。包括 、 和 在内的多能性基因在eMSCs中高表达。此外，培养的eMSCs对间充质干细胞标志物CD44、CD49f和CD105呈阳性，而对内皮细胞标志物CD31和造血细胞标志物CD34呈阴性。此外，未使用参考RNA测序来分析eMSCs的转录表达谱和关键途径。与皮肤成纤维细胞组相比，共鉴定出9104个差异表达基因（DEGs），其中5034个基因上调，4070个基因下调，基因本体论（GO）和京都基因与基因组百科全书（KEGG）中DEGs的前20个富集途径主要与G蛋白偶联受体信号通路、碳水化合物衍生物结合、核苷结合、核糖体生物发生、细胞周期、DNA复制、Ras信号通路和嘌呤代谢相关。在这些DEGs中，eMSCs组中一些促进间充质干细胞分化和增殖的代表性基因上调，而促进成纤维细胞增殖的基因下调。小熊猫eMSCs也具有多种分化能力，可分化为脂肪细胞、软骨细胞和肝细胞。总之，我们首次分离并鉴定了具有增殖和多向分化能力的小熊猫eMSCs。这种新的多能干细胞不仅有利于小熊猫种质资源的保护，也有利于未来的基础或临床前研究。
