Salient Achievements
Embryo genomics and Stem cell Physiology
An efficient technique was developed for the cryo-conservation of buffalo female
germplasm through rapid freezing with less volume of cryoprotectants in an indigenously
developed cryodevice, which yielded 100% post thaw recovery. Developed technology
showed that vitrification can improve, simplify and accelerate the cryopreservation
process of oocytes. Another important benefit of this technique is that no expensive
equipment is required. The technology developed did not have any adverse effect on
the intrinsic quality of the oocytes as the vitrification protocol employed had no
impact on the expression pattern of developmentally important genes of in vitro
matured oocytes. The vitrification protocol may be used for cryopreservation of
in vitro matured oocytes and this technology can be used for the farm species, it
has potential to be commercialized.
A novel and efficacious 3D microenvironment for the extended in vitro culture of
buffalo preantral follicles and embryonic stem cells was developed using collagen
gel surface matrix and gel droplet micro encapsulation.
Buffalo embryos were produced in vitro with 70-80% cleavage rate and 21% blastocysts.
To access the quality of these embryos, morphology, total cell count and expression
profiling of developmentally important genes was employed. These blastocysts were
used for the isolation of the inner cell mass for deriving embryonic stem cells.
ESCs were successfully grown over feeder layer and primary colonies could be
characterized with the specific markers for stem cells. Early stages of the ESCs
were Alkaline phosphatase positive and the stem cell clones showed the positive
staining for the marker proteins of nanog, Oct-4 and Sox-2. Stem cell clones have
been generated from early stage IVF derived embryos in goat and buffalo.
Parthenogenetic stem cell clones have been generated from early stage parthenogenetically
derived embryos in goat and buffalo.
Stress Physiology
As an ameliorative measure in buffaloes, extra electrolytes are essential during
heat stress to maintain electrolyte balance. Supplementation of NaHCO3 and K2CO3
help in maintaining blood Ph. Ascorbic acid in its polyphosphate form and zinc as
zinc oxide help in maintenance of homeostasis in stressed animals. A combination of
salt, ascorbic acid, zinc and probiotics (yeast) are good for lactating animals
during hot summer days to observe predicted effect on milk yield.