Establishment and Biological Characteristics of Fibroblasts from Different Origins in Asian Elephant
[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.
Asian elephantfibroblast cellsbiological characteristics
万方数据
维普
