亚洲象不同组织来源成纤维细胞系的建立及其生物学特性研究

doi:10.16431/j.cnki.1671-7236.2024.09.005

Abstract

Abstract: 【Objective】 The aim of this study was to establish Asian elephant fibroblast cell lines of different age and tissue source, and conduct cryopreservation and cell biological characteristics, so as to provide theoretical basis for the preservation and development of endangered animal genetic resources. 【Method】 Samples from 6-year-old Asian elephant ear edge tissue (AE00), newborn Asian elephant umbilical cord tissue (AE01), and 4-year-old Asian elephant ear edge tissue (AE02) 16 hours after death were collected, and fibroblast cell lines were established for passage culture and cryopreservation.The biological characteristics of fibroblasts from different age and tissue sources were analyzed by cell morphology, reproductive ability, aging, and apoptosis levels.Green fluorescent protein (GFP) transfection screening and karyotype detection were performed to analyze the genetic stability of Asian elephant fibroblasts. 【Result】 Three Asian elephant fibroblast cell lines were successfully established and cryopreserved.After recovery, the cells maintained a certain level of vitality.After continuous passage and culture, the cell morphology was spindle shaped.AE02 fibroblasts had the best cryopreservation activity and cell proliferation ability, with an apoptosis rate of AE01 (16.7%)＞AE00 (6.4%)＞AE02 (3.8%), and an aging rate of AE00 (35.9%)＞AE01 (17.2%)＞AE02 (7.8%).After 19 passages, the number of chromosomes in the AE02 fibroblasts remained stable.GFP transfection screening of AE00 resulted in stable expression of GFP in Asian elephant fibroblast cell lines. 【Conclusion】 In this study, Asian elephant fibroblast cell lines with different ages and tissue sources were successfully established.The cell morphology was spindle shaped, and the proliferation ability, cell aging, and apoptosis levels of fibroblast cell lines with different ages and tissue sources were different.After continuous passage, the number of cell chromosomes remained stable, enabling stable transfection and long-term screening of exogenous genes.It could be an important method for the long-term preservation and revival of endangered animal genetic resources.

Key words: Asian elephant; fibroblast cells; biological characteristics

CLC Number:

Q952

LI Jinda, PENG Ziwei, Muhammad Ameen Jamal, JIAO Deling, WEI Hongjiang. Establishment and Biological Characteristics of Fibroblasts from Different Origins in Asian Elephant[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3752-3761.

References

[1] NORRIS K,TERRY A,HANSFORD J P,et al.Biodiversity conservation and the earth system:Mind the gap [J].Trends in Ecology & Evolution,2020,35(10):919-926.
[2] HE C,DU J,ZHU D,ZHANG L.Population viability analysis of small population:A case study for Asian elephant in China [J].Integrative Zoology,2020,15(5):350-362.
[3] GHILDIYAL K,NAYAK S S,RAJAWAT D,et al.Genomic insights into the conservation of wild and domestic animal diversity:A review [J].Gene,2023,886:147719.
[4] YÁNEZ-ORTIZ I,CATALÁN J,RODRÍGUEZ-GIL J E,et al.Advances in sperm cryopreservation in farm animals:Cattle,horse,pig and sheep [J].Animal Reproduction Science,2022,246:106904.
[5] TAKEO T,NAKAO S,MIKODA N,et al.Optimized protocols for sperm cryopreservation and in vitro fertilization in the rat [J].Laboratory Animals (NY),2022,51(10):256-274.
[6] CHEN H,ZHANG L,MENG L,et al.Advantages of vitrification preservation in assisted reproduction and potential influences on imprinted genes [J].Clinical Epigenetics,2022,14(1):141.
[7] ZHENG Y L.Recent progress in reproduction of whale oocytes [J]. Zygote,2013,21(3):246-249.
[8] LAWRENZ B,COUGHLAN C,MELADO L,et al.The ART of frozen embryo transfer:Back to nature [J].Gynecological Endocrinology,2020,36(6):479-483.
[9] BORTOLETTO P,PRABHU M,BAKER V L.Association between programmed frozen embryo transfer and hypertensive disorders of pregnancy [J].Fertility and Sterility,2022,118(5):839-848.
[10] ECKER A,LÁZÁR B,TÓTH R I,et al.The effects of freezing media on the characteristics of male and female chicken primordial germ cell lines [J].Life (Basel),2023,13(4):867.
[11] KACZMARCZYK D,WOLNICKI J.Management of genetic variation in the Gamete Bank of the Endangered Lake Minnow Eupallasella percnurus,using Genassemblage 2.2 Software [J]. Animals (Basel),2022,12(23):3329.
[12] RODRIGUES L L V,MOURA Y B F,VIANA J V S,et al.Comparison between concentration and type of intracellular cryoprotectants and the presence of sucrose for cryobanks of somatic cells derived from captive Pumas [J].Zoo Biology,2023,42(3):440-448.
[13] MATHAN,ZAIB G,JIN K,et al.Formation,application,and significance of chicken primordial germ cells:A review [J].Animals (Basel),2023,13(6):1096.
[14] KENDALL R T,FEGHALI-BOSTWICK C A.Fibroblasts in fibrosis:Novel roles and mediators[J].Frontiers in Pharmacology,2014,5:123.
[15] BAI C,LI C,JIN D,et al.Establishment and characterization of a fibroblast line from landrace [J].Artificial Cells Nanomedicine & Biotechnology, 2010,38(3):129-135.
[16] LIU C,GUO Y,LU T,et al.Establishment and genetic characteristics analysis of in vitro culture a fibroblast cell line derived from Wuzhishan miniature pig [J]. Cryobiology,2014,68(2):281-287.
[17] TASRIPOO K,SUTHIKRAI W,SOPHON S,et al.First cloned swamp buffalo produced from adult ear fibroblast cell[J].Animal,2014,8(7):1139-1145.
[18] ELYASI GORJI Z,FARZANEH P,NASIMIAN A,et al.Cryopreservation of Iranian Markhoz goat fibroblast cells as an endangered national genetic resource [J].Molecular Biology Reports,2021,48(9):6241-6248.
[19] LIU Y,CHEN J,FENG T,et al.The establishment of giant panda (Ailuropoda melanoleuca) fibroblast cell line [J].In Vitro Cellular and Developmental Biology Animal,2022,58(3):194-198.
[20] UPHOFF C C,DREXLER H G.Comparative PCR analysis for detection of mycoplasma infections in continuous cell lines [J].In Vitro Cellular and Developmental Biology Animal,2002,38(2):79-85.
[21] HOLT W V,PICKARD A R.Role of reproductive technologies and genetic resource banks in animal conservation [J].Reviews of Reproduction,1999,4(3):143-150.
[22] ANDRABI S M,MAXWELL W M.A review on reproductive biotechnologies for conservation of endangered mammalian species [J].Animal Reproduction Science,2007,99(3-4):223-243.
[23] HANKS J.Conservation strategies for Africa’s large mammals [J].Reproduction Fertility and Development,2001,13(7-8):459-468.
[24] FAIR T,LONERGAN P,DINNYES A,et al.Ultrastructure of bovine blastocysts following cryopreservation:Effect of method of blastocyst production [J].Molecular Reproduction and Development,2001,58(2):186-195.
[25] WOODS E J,BENSON J D,AGCA Y,et al.Fundamental cryobiology of reproductive cells and tissues [J].Cryobiology,2004,48(2):146-156.
[26] SIENGDEE P,KLINHOM S,THITARAM C,et al.Isolation and culture of primary adult skin fibroblasts from the Asian elephant (Elephas maximus) [J]. PeerJ,2018，6:e4302.
[27] ELYASI GORJI Z,FARZANEH P,NASIMIAN A,et al.Cryopreservation of Iranian Markhoz goat fibroblast cells as an endangered national genetic resource [J].Molecular Biology Reports，2021，48(9):6241-6248.

Establishment and Biological Characteristics of Fibroblasts from Different Origins in Asian Elephant

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