Cell culture

The present study describes the first cell lines produced from members of class Chondrichthyes. Explants of brain tissue from Carcharhinus falciformis (silky shark) and Ginglymostoma cirratum (nurse shark) were incubated in a mammalian medium modified with the addition of urea, trimethylamine N-oxide, NaCl, and bovine serum. Primary monolayers were passaged with 0.025% trypsin in a modified saline solution. Silky shark cells grew optimally at 29C. The population doubling time for C. falciformis cells at passage 29 was 67 hours. For G. cirratum cells at passage 6 the population doubling was 84 hours. Silky shark cells grew over a broad range of osmolalities from 315 mOsm to a 1664 mOsm with optimal growth at 650 mOsm. A medium containing 10% dimethylsulfoxide allowed for cryopreservation with greater than 65% viability upon recovery. Current theories of elasmobranch osmoregulation are discussed in light of experimental data collected from studies conducted on the silky shark cell line.

Liver, kidney and spleen cells from healthy horses
and horses with equine infectious anemia were cultured
in vitro. Primary culture of organs from healthy horses
yielded typical elongate fibroblast cells. Primary culture
of organs from diseased horses yielded a variety of cell
types; pleomorphic giant cells being most common. The giant
cells did not divide, but either underwent degeneration and
death or became transformed into squamous-like cells lacking
contact inhibition resulting in the production of foci.
Inoculation of normal equine fibroblasts with either serum,
plasma, or liver, kidney or spleen extracts from diseased
horses resulted in a proliferation of the fibroblasts. Normal liver or kidney fibroblasts cocultivated with equine
leucocytes and inoculated with serum or plasma from a diseased
horse became transformed. This research supports the
proposal that equine infectious anemia virus is an oncornavirus.