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. MATERIAL AND 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%. CONCLUSION: 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.
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. MATERIAL AND 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%. CONCLUSION: 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: Yongpeng Gu; Hongzhen Li; Jun Miki; Kee-Hong Kim; Bungo Furusato; Isabell A Sesterhenn; Wei-Sing Chu; David G McLeod; Shiv Srivastava; Charles M Ewing; William B Isaacs; Johng S Rhim Journal: Exp Cell Res Date: 2006-01-17 Impact factor: 3.905