Abstract:To establish the caudal fin cell line of Schizothorax biddulphi, this study employed the tissue block method with fetal bovine serum (FBS) DME/F12 medium for in vitro cultivation of the tail fin, using previously established tail-fin cell lines (BICF1) of S. biddulphi. In this study, the effect of NaCl salinity and NaHCO3 alkalinity on the proliferation of tail fin cells was investigated using the MTT method. The main results are as follows: the BICF1 suspension culture was propagated to 45 generations. The optimum medium was DME/F12. The optimal FBS concentration was determined to be 20%. The optimum temperature was 25℃. The population doubling time of the 10th generation BICF1 was 28.11 h, showing an "S" type growth. The 6th generation BICF1 in liquid nitrogen was recovered after freezing for 180 days. Trypan blue staining showed that after recovery, (87.85±0.66)% of BICF1 cells were active, and the cells could proliferate and pass to next generation. The BICF1 cell line is free of contamination by bacteria, fungi, and mycoplasma. The sequencing results of mitochondrial 16S rRNA of the 10th generation BICF1 were consistent with the GenBank gene sequence, and the consistency rate of BICF1 and JQ844133.1 was 100%, proving that BICF1 was from S. biddulphi. BICF1 cell proliferation increased with salinity of 1, 2, 4, and 6, but decreased with salinity of 6, 8, and 10; BICF1 cell proliferation was the highest at 6. The NaHCO3 alkalinity of BICF1 increased at 2, 3, 4, 5, and 6 g/L, whereas the proliferation of BICF1 cells decreased at 6, 7, 8, and 10 g/L, respectively. BICF1 cell proliferation was the highest at 6 g/L. Cell proliferation first increased and then decreased with increasing salinity and alkalinity. This study aimed to provide a scientific basis for the rational development and utilization of genetic resources of S. biddulphi, including the establishment and protection of the germplasm.
