title: Establishment and biological characterization of fibroblast cell line from the Langshan chicken.
abstract: OBJECTIVE
  We needed to establish an embryonic fibroblast cell line from the Langshan chicken (LSCEF61) to preserve their important genetic resources at the cellular level.
  
  
  METHODS
  The cell line was established from 9-day-old embryos by direct explant culture and cryopreservation techniques. Cell morphology, dynamic proliferation and any contamination present were tested, and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analysed. Four types of fluorescent protein exogenous genes for pEGFP-C(1), pEGFP-N(3), pEYFP-N(1) and pDsRed1-N(1) were transfected into the cells.
  
  
  RESULTS
  Showed that the cells were healthy and were of spindle shaped structure, without change in morphology. Cell growth curves were of typical S-shape. Assays for microbial contamination were negative. The LSCEF61 line showed no cross-contamination when assessed by isoenzyme analysis. Chromosome number (2n) = 78 on more than 90% of occasions. The four types of fluorescent protein extro-genes appeared to be expressed effectively with high transfection efficiency between 15.6% and 38.6%.
  
  
  CONCLUSIONS
  The cell line met each of the quality control standards required for the American Type Culture Collection. It had not only preserved the genetic resources of the important Langshan chicken at the cellular level, but also provided valuable material for genomic, post-genomic and somatic cell cloning research and other applications.
authors.affiliation: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: W
authors.initials: W
authors.lastname: Guan
authors.forename: X
authors.initials: X
authors.lastname: He
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.forename: H
authors.initials: H
authors.lastname: Liang
authors.forename: Q
authors.initials: Q
authors.lastname: Zhao
authors.forename: Y
authors.initials: Y
authors.lastname: Pu
authors.forename: Y-H
authors.initials: YH
authors.lastname: Ma
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D002875:Chromosomes; D015925:Cryopreservation; D004171:Diploidy; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007621:Karyotyping; D045022:Lactate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D009924:Organ Culture Techniques; D011786:Quality Control; D014162:Transfection
chemical_list: D007527:Isoenzymes; D045022:Lactate Dehydrogenases; D008291:Malate Dehydrogenase
references.citation: Exp Cell Res. 2006 Apr 1;312(6):831-43
references.pmid: 16413016
references.citation: Vet Immunol Immunopathol. 2006 Sep 15;113(1-2):30-6
references.pmid: 16678911
references.citation: Brain Res. 2002 Nov 29;956(2):221-9
references.pmid: 12445689
references.citation: J Cell Physiol. 2007 Jan;210(1):63-71
references.pmid: 17013808
references.citation: Int J Cancer. 1999 Nov 12;83(4):555-63
references.pmid: 10508494
references.citation: Biochem Biophys Res Commun. 2006 Jul 7;345(3):926-32
references.pmid: 16707099
references.citation: In Vitro Cell Dev Biol Anim. 1998 Jan;34(1):35-9
references.pmid: 9542633
references.citation: Vet Immunol Immunopathol. 2004 Jan;97(1-2):65-76
references.pmid: 14700538