From endangered to preserved: Biobanking of the Katchaikatti Black sheep through dermal fibroblast cryopreservation
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Keywords:
Cryopreservation, Endangered, Karyotyping, Katchaikatti Black sheep, Skin fibroblast cellsAbstract
The present study was carried out with the aim of ex-situ conservation of the elite germplasm of Indian sheep
breeds including Katchaikatti Black sheep in the form of cryo cell bank repository. The Katchaikatti Black sheep
are distributed in the villages of Vadipatti block of Madurai district in Southern Tamil Nadu and belong to the
meat type sheep with extremely coarse and hairy wool. As on date, Katchaikatti black sheep breed is at the risk
of extinction and considered to be endangered. It is therefore imperative to conserve the highly valuable sheep
germplasm of Indian origin. In this connection, as a preliminary step, a total number of eight ear pinna samples
were collected from eight individual sheep (four males and four females from Katchaikatti village) using the sterile
surgical blade, washed once in 70 % alcohol, saturated once in washing medium, stored in complete cell culture
medium and transported under cold chain. Once in six hours, the complete medium was changed. After bringing
to the laboratory at ICAR-CSWRI, Avikanagar, standard cell culture protocol was followed for the initial tissue
explant culture. Subculturing of the resultant somatic fibroblast cell was done up to the passage four level at the
periodical intervals. In order to use the skin fibroblasts in downstream processing, karyotyping was also carried
out through outsourcing, as a gold standard. Various parameters such as viability assay, population doubling
time were also carried out. As a final step, cryopreservation of the skin fibroblast cells derived from Katchaikatti
black sheep was done using LN2 protocol, after the completion of the standard characterization regimen.
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