承德无角山羊耳成纤维细胞系的建立与冷冻保存研究

doi:10.16431/j.cnki.1671-7236.2024.01.019

Abstract

Abstract: 【Objective】 In this study, betaine was used as a cryoprotectant to cryopreserve Chengde Hornless goat ear fibroblast cell lines for optimizing the cryopreservation effect, so as to preserve the genetic resources of this local breed at the cellular level.【Method】 The ear fibroblast cell line of Chengde Hornless goat was established by tissue block culture method.Vitrification of fibroblasts with betaine as a cryoprotectant, different concentrations of betaine (5%, 10%, 15%, 20% and 25%) were used as the experimental group, and different concentrations of dimethyl sulfoxide (DMSO, 10% and 20%) were used as the control group.Cryopreservation of Chengde Hornless goat ear fibroblasts was performed, the cell survival rate, cell proliferative activity and the expression of B-cell lymphoma-2(Bcl-2)/Bcl-2 related X protein (Bax) genes were measured after cell resuscitation, and the toxicity of betaine was also assessed.【Result】 Chengde Hornless goat ear fibroblast cell line established in this study had good growth trend and high cell purity.The result of bacteria, fungi and mycoplasma were negative, and the growth curve was S type, which met the requirements of establishing fibroblast cell line.When the concentration of betain was 15%, the survival rate of cells was the highest, reaching 90.86%, which was extremely significantly higher than that of control group (P<0.01).The cell proliferation activity and the expression of Bcl-2/Bax genes were significantly higher than those in control group (P<0.05), and the toxicity of betain was much lower than that of DMSO (P<0.05).【Conclusion】 In this study, betaine was successfully used as a cryoprotectant to cryopreserve the ear fibroblast cell line of Chengde Hornless goat, so as to protect the genetic resources of Chengde Hornless goat.

Key words: Chengde Hornless goat; fibroblast cell line; cryopreservation; betaine

CLC Number:

S815

SUN Qingyi, LI Mengqi, ZHAO Wei, ZHANG Wentao, ZHANG Xiaosheng, ZHANG Jinlong, ZHANG Hongjun, FANG Xiaohuan, TAO Chenyu, XIA Wei, LI Junjie. Establishment and Cryopreservation of Chengde Hornless Goat Ear Fibroblast Cell Line[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 183-192.

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Establishment and Cryopreservation of Chengde Hornless Goat Ear Fibroblast Cell Line

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