executive summary - National Dairy Research Institute

Research Achievements
the expression level of THY1 in SSC-like cells is not
affected by GDNF, EGF and FGF2, together or in
combination and vi) GDNF increases whereas EGF
and FGF2 decrease the expression level of ETV5 in
Sertoli cells.
Production of Goat induced Pluripotent Stem (iPS)
Cells
Goat iPS cells were produced by transfecting Nanog,
Oct4, Sox2 and Lin28 to goat adult fibroblasts cells
using lentiviruses. Characterization of iPS cells was
carried out by examining expression of alkaline
phosphatase, intracellular markers Oct4, Nanog and
Sox2 and, surface markers SSEA-1, SSEA-3, SSEA-4,
TRA-1-60 and TRA-1-81. For directed differentiation
of iPS cells to germ cell-like cells, embryoid bodies
produced from iPS cells were cultured in the presence
of retinoic acid and BMP-4. The germ cell-like cells
generated were characterized by examining the
expression of germ cell markers such as VASA,
STELLA and PUM1 by immunofluorescence staining
and Western blotting.
Isolation, Characterization and Differentiation of
Goat Mesenchymal Stem Cells (MSCs)
MSCs were isolated from goat adipose tissue and were
subjected to characterization and directed
differentiation to different lineages. The Putative
MSCs were examined for the expression of surface
markers CD29, CD44, CD166, CD99 and CD34 by RT-
PCR. Both fresh and cryopreserved MSCs were
cultured to induce osteogenic, chondrogenic,
adipogenic andneurogenic differentiation, which were
characterized with their specific markers. Tests were
performed using Alzarine dye, surface marker
osteocalcin and collagen. MSCs were found to
differentiate into osteogenic cell lineages, which were
further confirmed by RT-PCR analysis for osteocalcin,
an osteocyte marker.
Buffalo LIF (BuLIF) was cloned, sequenced and
characterized for the first time and expressed in COS-1
cell line for its stable expression. The BuLIF ORF is
made of 609 nucleotides, which codes for 202 amino
acids long peptide. The initial 22 amino acids represent
signal peptide. The overall similarity of BuLIF with
bovine, human and mouse is 92.00%, 83.38% and
72.74% at nucleic acids level and 98.52%, 89.66% and
77.45% at amino acids level respectively.
Bioinformatics analysis indicates BuLIF to be a
Annual Report - 2012-13
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