Cloned buffalo `Garima-II` born at Karnal

The National Dairy Research Institute at Karnal has cloned a buffalo calf.

Updated: Aug 22, 2010, 23:33 PM IST

Chandigarh: 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 A K 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 new born 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 M S Chauhan, S K Singla, R S 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

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

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.