Establishment and characterization of Caspian horse fibroblast cell bank in Iran

Abstract

Caspian horse, a rare horse breed found in 1965 by Louise Firouz in northern Iran, is a small horse which is reported to be in danger of extinction in its original homeland. There seems to be a great need to prevent extinction of this valuable horse. In this study, 51 fibroblast cell lines from Caspian horse ear marginal tissue were successfully established by sampling 60 horses using primary explant technique. Cells were authenticated and growth curve was plotted. According to results obtained, population doubling time (PDT) was calculated 23 ± 0.5 h for all cell lines. Multiplex polymerase chain reaction (multiplex PCR) revealed that cell lines had no cross-contamination with other species. Bacteria, fungi, and mycoplasma contamination were checked using standard methods such as PCR, direct culture, and Hoechst staining. In addition to providing a valuable source for genomic, postgenomic, and somatic cloning researches, the established cell lines would preserve Caspian horse genetic resources. It will also create an accessible database for researchers.

Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines

“…These characteristics of explants during in vitro culture were similar to the explants derived from other domestic and wild mammals. In studies using tissues from horses, the migration of fibroblast and epithelial-like cells from explants have been observed after 5-7 days of an in vitro culture (Amoli et al, 2017). In the case of goat-derived tissues, the explants reportedly adhered to the flasks within 5-7 days and the cells became confluent within 3-5 days post adhesion (Bai et al, 2012).…”

“…For a morphological confirmation, the cells were subjected to an immunocytochemistry protocol based on the method described by Amoli et al (2017). Briefly, the cells were fixed using 4% paraformaldehyde for 10 min at 25 C, then washed with chilled PBS.…”

Peer Review #4 of "Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines (v0.1)"

“…These characteristics of explants during in vitro culture were similar to the explants derived from other domestic and wild mammals. In studies using tissues from horses, the migration of fibroblast and epithelial-like cells from explants have been observed after 5 days to 7 days of an in vitro culture (Amoli et al, 2017). In the case of goat-derived tissues, the explants reportedly adhered to the flasks within 5 days to 7 days and the cells became confluent within 3 days to 5 days post adhesion (Bai et al, 2012).…”

Peer Review #1 of "Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines (v0.2)"

“…Manuscript to be reviewed For a morphological confirmation, the cells were subjected to an immunocytochemistry protocol based on the method described by Amoli et al (2017). Briefly, the cells were fixed using 4% paraformaldehyde for min at 25°C, then washed with chilled PBS.…”