Somatic cell conservation of the Punganur cattle: An Indian heritage breed

Abstract

ICAR-NBAGR is the nodal agency for conserving indigenous livestock genetic resources as embedded in the
Sustainable Developmental Goal of the United Nations specifically 2.5.1.a. Indigenous Punganur cattle is one of
the shortest cattle breeds of the world that has <10,000 true to the breed animals (19th Livestock Census). Breed
Watch list 2022- ICAR-NBAGR, Karnal enlists Punganur as a vulnerable breed and hence it was considered on
priority for conservation. Ear skin tissue samples of both sexes were used for generating the somatic cells by
in vitro explant and cell culture. Initially, tissue fragments were cultured in Dulbecco’s Modified Eagle Medium
supplemented with 20% FBS and 2% antibiotic–antimycotic solution under a controlled atmosphere (37 o
C, 5%

CO2
). Fibroblast cells having fusiform shape and centrally positioned oval nuclei were isolated and multiplied
in culture till the 4th passage. The cell lines demonstrated stable growth characteristics and maintained normal
morphological features. At least 30 cryovials per animal (1x106 cells/ vial) were cryopreserved in a liquid nitrogen
container at -196 o

C for posterity. The growth potential of cryopreserved cells was found to be comparable to
that of the cells before freezing. Advances in the cloning of animals using somatic cells for potential revival of
a breed and future breakthroughs are likely to enhance the use of these established cell lines in diverse fields,
such as disease modeling and treatment, tissue engineering, cellular reprogramming, and the production of
extracellular matrix. Therefore, the stored biological material is valuable both for ex-situ conservation of the
genetic resources and for future research avenues.