Construction and Characterization of Immortalized Fibroblast Cell Line from Bactrian Camel
Author:
Yan Meilin1, Yong Fang1, Ji Wangye1, Zhang Lili1, Zhao Shuqin12, Gao Yuan12ORCID
Affiliation:
1. College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China 2. Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
Abstract
Immortalized cell lines with many advantages are widely used in various experimental contexts by many different labs. However, the absence of available cell lines poses difficulties for research in some species, such as camels. To establish an immortalized Bactrian camel fibroblast (iBCF) cell line and understand its biological characteristics, primary fibroblast cells from Bactrian camels were isolated and purified using enzymatic digestion in this study, and telomerase reverse transcriptase (hTERT) vectors were introduced into primary BCF (pBCF) for continuous passage to 80 generations after screening with G418. The cell morphology of different generations was examined under a microscope. Cell cycle and viability were evaluated by flow cytometry and CCK-8 assay, respectively. Cellular genes expression was monitored by qPCR, immunofluorescence, and Western blot, respectively. Chromosomes were determined by karyotyping. The results showed that like most other cells, both pBCF and iBCF were sensitive to nutrient concentrations and adapted to culture in the medium with 4.5 g/L glucose and 10% fetal bovine serum (FBS) concentration. hTERT gene was introduced and stably expressed in iBCF cells, which promoted BCF cell immortalization. The fibroblast specific marker vimentin (VIM) is expressed in both pBCF and iBCF, but epithelial marker cytokeratin18 (CK18) expression is weak in BCF cells. Proliferation and viability detection showed that hTERT-induced iBCF exhibits faster growth rates and higher viability than pBCF. Karyotyping showed that iBCF maintained the same number and morphology of chromosomes as the pBCF. This study demonstrated that we have successfully constructed an immortalized Bactrian camel fibroblast cell line, which was named BCF23. The establishment of the BCF23 cell line provides a foundation for expanding camel-related research.
Funder
the National Natural Science Foundation of China the Longyuan Youth Innovation and Entrepreneurship Talent Project
Subject
Paleontology,Space and Planetary Science,General Biochemistry, Genetics and Molecular Biology,Ecology, Evolution, Behavior and Systematics
Reference33 articles.
1. Gao, Y., Zhao, S., Zhang, W., Tang, H., Yan, M., Yong, F., Bai, X., Wu, X., Zhang, Y., and Zhang, Q. (2023). Localization of FGF21 Protein and Lipid Metabolism-Related Genes in Camels. Life, 13. 2. Chen, Y., Hu, S., Wang, M., Zhao, B., Yang, N., Li, J., Liu, M., Zhou, J., Bao, G., and Wu, X. (2019). Characterization and Establishment of an Immortalized Rabbit Melanocyte Cell Line Using the SV40 Large T Antigen. Int. J. Mol. Sci., 20. 3. Establishment and characterization of five immortalized human scalp dermal papilla cell lines;Kwack;Biochem. Biophys. Res. Commun.,2018 4. Establishment and characterization of immortalized erythroid progenitor cell lines derived from a common cell source;Kurita;Exp. Hematol.,2019 5. Establishment, characterization, and toxicological application of a spontaneous immortalized cell line from the striped field mouse, Apodemus agrarius;Lee;Vitr. Cell Dev. Biol. Anim.,2018
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