Cloned Buffalo "Garima -2 " born at NDRI,Karnal
Dr A K Srivastava said that this cloned buffalo calf was different from the earlier clone calf because in this case the used donor cell was embryonic stem cell. However, in earlier cloning, the donor cell was from somatic cells.
The donor embryonic stem cell was isolated from the 8 day old blastocyst. These cells were cultured up to 29-passages (117 days) till it expressed pluripotent marker and then confirmed to be stem cell.
Dr A K Srivastava, director, NDRI,emphasised that this technology could go a long way in helping faster multiplication of superior milch buffaloes in India. He said that although the world's largest population of buffaloes was in India and they were contributing about 55% of total milk production in the country, but the percentage of elite animals was very low and there was an urgent need to enhance the population of these elite buffaloes.
He further emphasised that there was an acute shortage of good bulls and the technology of cloning would decrease this gap between supply and demand of breeding the bulls in the shortest possible time. The team of the jubilant scientists involved in the production of this cloned calf using embryonic stem-cell as donor cell were Dr M S Chauhan, Dr S K Singla, Dr R S Manik, Dr P Palta, Dr Shiv Parsad, and Dr Aman George of NDRI, Karnal.
The scientists were of the opinion that the embryonic stem cells had better cloning ability as compared to somatic cells, as such the epigenetic reprogramming of these cells was much more efficient than the somatic cells, which were already differentiated and lineage committed.
Earlier NDRI had produced the world cloned buffalo calf on February 6, 2009.The hand-guided cloning technique developed at NDRI, is an advanced modification of the "Conventional Cloning Technique".
In this technique, immature oocytes were isolated from ovaries and were matured in vitro. These were then denuded and treated with an enzyme to digest the outer layer of oocytes called ‘zona pellucida’. The oocytes were then treated with chemicals to push their genetic material to one side of the oocyte.
This protruded side was then cut off with the help of "hand held fine blade" for removing the original genetic material of the oocyte.
The enucleated oocyte was then electro-fused with a single cell taken from a colony of embryonic stem cells. The resulting embryos were cultured and grown in the laboratory for seven days to develop them to the stage of blastocyst.The blastocysts were transferred to recipient buffaloes