More than a year after it successfully managed to clone a buffalo calf, the National Dairy Research Institute at Karnal has repeated the feat with the birth of a second calf.
The bovine was cloned using the advanced hand-guided technique, Dr AK Srivastava, Director, NDRI, Karnal, said.
"This cloned buffalo calf is different from the earlier cloned calf because in this case the used donor cell was an embryonic stem cell. In the earlier cloning, the donor cell was from somatic cells," he said, adding the newborn weighed 32 kg.
On February 6 last year, NDRI had produced the world's first cloned buffalo calf.
"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," said an NDRI spokesman quoting Srivastava.
Srivastava emphasised that this technology could go a long way in helping for faster multiplication of superior milch buffaloes in India.
He said although the world's largest population of buffaloes is in India and they are contributing about 55 per cent of total milk production in the country, the percentage of the animal is very less and there is an urgent need to enhance the population of these buffaloes.
He further said there is an acute shortage of bulls and cloning can decrease the gap between supply and demand of bulls in the shortest possible time.
The team of scientists involved in the production of this cloned calf using embryonic stem-cell as donor cell include MS Chauhan, SK Singla, RS Manik, P Palta, Shiv Parsad, and Aman George of NDRI.
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, the spokesman said.