title: Establishment and genetic characteristics analysis of in vitro culture a fibroblast cell line derived from Wuzhishan miniature pig.
abstract: Establishment of fibroblast cell lines of endangered pig breeds and research on the gene functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. The Wuzhishan miniature pig ear marginal tissue fibroblast cell line (WPF22) from 22 samples, stocking 87 cryogenically-preserved vials, was successfully established by using primary explants technique and cell cryopreservation techniques. WPF22 cells were adherent, with a population doubling time of 30.2h. Chromosome karyotyping and G-banding analysis showed that >90.2% of cells were diploid (2n=38) prior to the 4th generation. Neither microbial contamination nor cross-contamination was detected by isoenzyme analyses. Cell viability was 97.8% before cryopreservation and 94.9% after recovery. To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, six fluorescent protein genes were transferred into fibroblasts by lipofectamine-mediated method. The transfection efficiency of six fluorescent protein genes fluctuated between 8.1% and 42.6%. ECFP and DsRed were mostly shown in cytoplasmic in dots around the nucleus, and EYFP and EGFP had a slightly stronger expression in the nucleus than in the cytoplasm, but without expression in some vacuoles. Every index of the WPF22 cell line meets all the standard quality controls of American type Culture Collection (ATCC). This research thus does not only preserve important genetic resources of Wuzhishan miniature pig at the cell level, but also serve as a valuable resource for genome, postgenome and somacloning research.
authors.affiliation: Bioscience Department, Department of Laboratory Medicine, Bengbu Medical College, Bengbu 233030, China; Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100194, China.
authors.forename: Changqing
authors.initials: C
authors.lastname: Liu
authors.affiliation: Bioscience Department, Department of Laboratory Medicine, Bengbu Medical College, Bengbu 233030, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100194, China.
authors.forename: Taofeng
authors.initials: T
authors.lastname: Lu
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100194, China.
authors.forename: Xiangchen
authors.initials: X
authors.lastname: Li
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100194, China. Electronic address: wjguan86@hotmail.com.
authors.forename: Weijun
authors.initials: W
authors.lastname: Guan
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100194, China.
authors.forename: Yuihui
authors.initials: Y
authors.lastname: Ma
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D056727:Endangered Species; D005347:Fibroblasts / Q000166:cytology*; D013552:Swine; D013556:Swine, Miniature
keywords: Characterization; Cryopreservation; Fibroblast cell line; Wuzhishan miniature pig