Preliminary study on the establishment of kidney tissue cell lines in Schizothorax eurystomus

Abstract:

Fish cell culture is an important model for germplasm preservation, gene function analysis, and cell engineering breeding. Therefore, it is of great significance to establish cell lines for the endangered Schizothorax eurystomus. The aim of this study was to establish a kidney tissue cell line for S. eurystomus, for which mesonephric tissue was used. The primary cell culture was initiated by tissue block transplantation and the subculture was carried out after the cells were stabilized. A kidney cell line (EUM10) was established. To analyze the cryopreservation and resuscitation ability of renal cell lines and the optimal growth conditions for S. eurystomus, PCR was used to detect contamination and mitochondrial gene fragment analysis was used to identify the source of the cell. The results showed that the optimal medium for S. eurystomus kidney tissue cells was DME/F-12, the optimal temperature was 25℃, and the optimal serum concentration was 20%. The cells grew in suspension and the growth curve was S-shaped. The growth curve shows that the population doubling time of the kidney tissue cell line cells in the 10th generation was 23.26 h. After 6 months of cryopreservation with liquid nitrogen, the recovery/survival rate was 91.91% (6th generation). S. eurystomus shares 99.81% of the gene sequence identity with a split-bellied fish from Tarim, which proves that the cell line originates from a wide-mouthed split-bellied fish. In this study, a kidney cell line for S. eurystomus was successfully established. Kidney cell lines can be used as in vitro systems to study various aspects of fish, such as cytogenetics and gene function analysis. This study provides the basic data for the initial research into and breeding of S.eurystomus.