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Characteristics of mesenchymal stem cells and their exosomes derived from giant panda (Ailuropoda melanoleuca) endometrium

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Abstract

Conservation of genetic resources is an important way to protect endangered species. At present, mesenchymal stem cells (MSCs) have been isolated from the bone marrow and umbilical cords of giant pandas. However, the types and quantities of preserved cell resources were rare and limited, and none of MSCs was derived from female reproductive organs. Here, we first isolated MSCs from the endometrium of giant panda. These cells showed fibroblast morphology and expressed Sox2, Klf4, Thy1, CD73, CD105, CD44, CD49f, and CD105. Endometrium mesenchymal stem cells (eMSCs) of giant panda could induce differentiation into three germ layers in vitro. RNA-seq analysis showed that 833 genes were upregulated and 716 genes were downregulated in eMSCs compared with skin fibroblast cells. The results of GO and the KEGG analysis of differentially expressed genes (DEGs) were mainly focused on transporter activity, signal transducer activity, pathways regulating pluripotency of stem cells, MAPK signaling pathway, and PI3K-Akt signaling pathway. The genes PLCG2, FRK, JAK3, LYN, PIK3CB, JAK2, CBLB, and MET were identified as hub genes by PPI network analysis. In addition, the exosomes of eMSCs were also isolated and identified. The average diameter of exosomes was 74.26 ± 13.75 nm and highly expressed TSG101 and CD9 but did not express CALNEXIN. A total of 277 miRNAs were detected in the exosomes; the highest expression of miRNA was the has-miR-21-5p. A total of 14461 target genes of the whole miRNAs were predicted and proceeded with functional analysis. In conclusion, we successfully isolated and characterized the giant panda eMSCs and their exosomes, and analyzed their functions through bioinformatics techniques. It not only enriched the conservation types of giant panda cell resources and promoted the protection of genetic diversity, but also laid a foundation for the application of eMSCs and exosomes in the disease treatment of giant pandas.

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Funding

This work was supported by the Natural Science Foundation of Sichuan Province (2022NSFSC1608 and 2023NSFSC0170), Chengdu Giant Panda Breeding Research Foundation (CPF2017–16), and Chengdu Research Base of Giant Panda Breeding (2022CPB-B11).

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Authors and Affiliations

  1. Chengdu Research Base of Giant Panda Breeding, Sichuan Province, Chengdu, 610081, China

    Yu-Liang Liu, Jia-Song Chen, Jun-Hui An, Zhi-Gang Cai, Jing-Chao Lan, Xiang-Wei Kong, Ming-Yue Zhang, Rong Hou & Dong-Hui Wang

  2. Sichuan Key Laboratory of Conservation Biology for Endangered Wildlife, Sichuan Province, Chengdu, 610081, China

    Yu-Liang Liu, Jia-Song Chen, Jun-Hui An, Zhi-Gang Cai, Jing-Chao Lan, Yuan Li, Xiang-Wei Kong, Ming-Yue Zhang, Rong Hou & Dong-Hui Wang

  3. Sichuan Academy of Giant Panda, Sichuan Province, Chengdu, 610081, China

    Yu-Liang Liu, Jia-Song Chen, Jun-Hui An, Zhi-Gang Cai, Jing-Chao Lan, Xiang-Wei Kong, Ming-Yue Zhang, Rong Hou & Dong-Hui Wang

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  1. Yu-Liang Liu
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  2. Jia-Song Chen
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  3. Jun-Hui An
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  6. Yuan Li
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  8. Ming-Yue Zhang
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  9. Rong Hou
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  10. Dong-Hui Wang
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Contributions

Yu-Liang Liu: conceptualization, methodology. Jia-Song Chen: visualization, investigation. Jun-Hui An: formal analysis. Zhi-Gang Cai: formal analysis. Jing-Chao Lan: visualization, investigation. Yuan Li: investigation. Xiang-Wei Kong: investigation. Ming-Yue Zhang: formal analysis. Rong Hou: project administration. Dong-Hui Wang: conceptualization, methodology, writing-original draft, writing-review and editing. All the authors have contributed to this study and are aware of submission.

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Correspondence to Dong-Hui Wang.

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Lay summary

We for the first time successfully isolated and characterized the giant panda endometrial mesenchymal stem cells (eMSCs) and their exosomes, and analyzed their functions through bioinformatics techniques. It not only enriched the conservation types of giant panda cell resources but also laid a foundation for disease treatment of giant pandas.

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Liu, YL., Chen, JS., An, JH. et al. Characteristics of mesenchymal stem cells and their exosomes derived from giant panda (Ailuropoda melanoleuca) endometrium. In Vitro Cell.Dev.Biol.-Animal 59, 550–563 (2023). https://doi.org/10.1007/s11626-023-00802-1

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