Abstract :
An ear marginal fibroblast cell bank was established from the Jining Black Grey (JBG) goat
using attachment culture and freezing biotechniques. This bank included 32 ear samples
(15 males and 17 females) and has stocks of 168 cryogenically preserved vials, each vial
contained 4.0×106 cells per milliliter. The cells of the bank that were checked for the
quality and the biological characteristics showed a typical fibroblast morphology when
they cultured in vitro. The growth curve consisted of a growth curve consisting of a latent
phase, logarithmic growth phase and stationary phase, cell population doubling time (PDT)
of 48 h. The chromosome analysis showed that the frequency of cells having the diploid
number of chromosomes (60) was 98.65±2.89%, and no microbe contamination (bacteria,
epiphyte, virus or mycoplasma) was detected. In addition, lactate dehydrogenase (LDH) and
malate dehydrogenase (MDH) zymography indicated that this cell bank was free of crosscontamination.
At 24, 48 and 72 h after transfection, the expression efficiency of pEGFP-C1,
pEGFP-N3, pEYFP-N1, pECFP-N1, pECFP-mito and pDsRed1-N1 were between 11.8% and
56.3%. The fluorescence could be observed well-distributed in cytoplasm and nucleus except
for some cryptomere vesicles at 24 h after transfection. These newly established cell lines
meet all the quality control standards established by the American Type Culture Collection.
We have employed a new method for conserving the genetic resources of an important and
endangered animal breed. The fibroblast bank that we have established from the JBG goat
also provides an invaluable material resource for future studies that will utilize molecular
and cell biology applications.
