TY - JOUR TI - Establishment and characterization of fibroblast cultures derived from a female common hippopotamus (Hippopotamus amphibius) skin biopsy AU - Wang, Tao AU - Li, Zelong AU - Wei, Jinpu AU - Zheng, Dongmin AU - Wang, Chen AU - Xu, Chang AU - Chen, Wu AU - Wang, Bo T2 - Cell Biology International AB - The population decline of the common hippopotamus (Hippopotamus amphibius) has necessitated the preservation of their genetic resources for species conservation and research. Of all actions, cryopreservation of fibroblast cell cultures derived from animal biopsy is considered a simple but efficient means. Nevertheless, preserving viable cell cultures of the common hippopotamus has not been achieved to our knowledge. To this end, we established and characterized fibroblast cell cultures from the skin sample of a newborn common hippopotamus in this study. By combining the tissue explant direct culture and enzymatic digestion methods, we isolated a great number of cells with typical fibroblastic morphology and high viability. Neither bacteria/fungi nor mycoplasma was detectable in the cell cultures throughout the study. The population doubling time was 34 h according to the growth curve. Karyotyping based on Giemsa staining showed that the cultured cells were diploid with 36 chromosomes in all, one pair of which was sex chromosomes. Amplified mitochondrial cytochrome C oxidase subunit I gene sequence of the cultured cells was 99.26% identical with that of the registered Hippopotamus amphibius complete mitochondrial DNA, confirming the species of origin of the cells. Flow cytometry and immunofluorescence staining results revealed that the detected cells were positive for fibroblast markers, S100A4 and Vimentin. In conclusion, we generated the fibroblast cell cultures from a common hippopotamus and identified their characteristics using multiple techniques. We believe the cryopreserved cells could be useful genetic materials for future research. This article is protected by copyright. All rights reserved. DO - https://doi.org/10.1002/cbin.11596 DP - Wiley Online Library VL - n/a IS - n/a LA - en SN - 1095-8355 UR - https://onlinelibrary.wiley.com/doi/abs/10.1002/cbin.11596 Y2 - 2021/03/29/23:25:56 KW - Characteristics KW - Common hippopotamus KW - Cryopreservation KW - Fibroblast cell culture KW - Isolation KW - Skin sample ER -