Cell lines have been established to preserve the genetic material of endangered
animals. This study aims to establish, characterize and authenticate fibroblast cells derived
from the kidney tissue of a Sumatran rhinoceros’ carcass. The primary cultures were
obtained using the mixed enzymatic-explant method, supplemented with complete media
and maintained at 37ºC with 5% CO2
in an incubator. Following routine trypsinization,
viability and growth curves were generated through the Trypan Blue counting method.
Cellular senescence was quantified by Sa-β-gal staining assay and G-banding for
karyotyping. As a result, the cell derivation had generated 81 frozen stocks. The viability of
cells at P5 and P10 showed reasonable recovery after six months. Cell population doubling
time at P5 was 20.45 hours, while it was 22.35 hours at P10 and P15. The senescence
level significantly increased from P5 to P10, and was especially significant at P15.
Genetic stabilities were considered stable at P5 and P10, with frequency of over 70 %. In
conclusion, this study was able to derive a primary fibroblast culture from the preserved
tissue of a Sumatran rhinoceros, with certain changes in morphology, senescence level,
growth curves and cell viability as the number of passages increased. |
