Impact of uterine epithelial cells and its conditioned medium on the in vitro embryo production in buffalo (Bubalusbubalis).

One of the challenges usually needs to be addressed in animal embryo production is to create the appropriate in vitro culture to improve blastocyst rate and produce high quality embryos. The present work was undertaken to investigate the impact of uterine epithelial cells and its conditioned medium (CM) on in vitro embryo production in buffalo. Buffalo uterine epithelial cells (UECs) were successfully isolated, cultured and characterized from slaughterhouse derived non gravid uteri.

Effects of Growth Factors on Establishment and Propagation of Embryonic Stem Cells from Very Early Stage IVF Embryos and Their Characterization in Buffalo.

Background And Objectives: Although ES cells have been derived from very early stage embryos in different species, but, so far ES cell line could be derived from early stage IVF embryos in buffalo. The present experiment was carried out to study the effects of different growth factors on attachment, formation of ES cell colonies, their extent of passaging and relative expression of pluripotency marker in these colonies in buffalo.

Methods And Results: For this, 8~16 cell stages zona free IVF embryos were cultured with different culture condition viz.

Comparative expression analysis of embryonic development-related genes at different stages of parthenogenetic and in vitro fertilized embryos in caprine.

Aberrant gene expression occurs in parthenogenetic embryos due to abnormal epigenetic modifications in the genome that probably diminish viability and enhance developmental abnormalities in these embryos. In the present study, five developmentally important genes (HPRT1, Cx43, Sox2, Mest and IGF2R) were analysed at different stages in parthenotes (haploid and diploid) and compared with similar stages in in vitro fertilized (IVF) embryos. The results indicated that in haploid parthenotes expression of HPRT1 was upregulated (P < 0.

Generation of transgenic mesenchymal stem cells expressing green fluorescent protein as reporter gene using no viral vector in caprine.

Mesenchymal stromal cells (MSC) are multipotent cells that can be derived from many different organs and tissues. While there are many ways to label and track cells each with strengths and weakness, the green fluorescent protein (GFP) is a reporter gene commonly employed. In the present study, caprine MSC were collected from bone marrow and cells were characterised with MSC specific markers.

Effect of actin polymerization inhibitor during oocyte maturation on parthenogenetic embryo development and ploidy in Capra hircus.

This study was designed to observe the effect of cytochalasin B (CCB) concentrations on ploidy and early development of parthenogenetic embryos in a caprine species. Caprine oocytes were matured in the presence of different concentrations of CCB (5, 10, 15, and 20 μg/ml) and activated by 7% ethanol followed by incubation with 2 mM DMAP. For embryos fertilized in vitro, oocytes were matured in maturation medium without CCB.

Reprogramming of fetal cells by avian EE for generation of pluripotent stem cell like cells in caprine.

The present work was carried out to study the ability of avian "Extract Egg" (EE) for reprogramming caprine fetal cells. The isolated caprine fetal cells were cultured in stem cell media supplemented with different percentages of either EE or FBS. The results indicated that the supplementation of 2-4% EE formed lesser but larger size stem cell like cell colonies as compared to 6% or 10% EE.

Survivability of parthenogenetic embryos following in vivo transfer in naturally synchronized Capra hircus.

The present study was conducted to see the in vivo developmental potency of caprine parthenogenetic embryos generated in a modified way. The good quality caprine oocytes were matured in presence of cytochalasin B (CCB) and then activated by 7% ethanol followed by 2 mM 6-dimethyl amino purine (6-DMAP) and embryo development was recorded. Early stage parthenogenetic embryos (two to four cells) were surgically transferred in recipients (10).

Effect of mitotic inducers and retinoic acid blocker on expression of pluripotent genes in ES cells derived from early stage in vitro-produced embryos in buffalo.

So far, it has been difficult to generate embryonic stem (ES) cell from early stage preimplantation embryos of buffalo. These ES cells will be more helpful for efficient embryo cloning and generation of body cells as they are more primitive than inner cell mass (ICM)-derived ES cells. The present study was conducted to find the effect of lipopolysaccharide (LPS), melatonin (N-acetyl-5-methoxytryptamine, a pineal gland product), and citral (3,7-dimethyl-2,6-octadienal and a retinoic acid synthesis blocker) on establishment of primary ES cell colonies, the comparative size of the ES cell colonies, and expression of pluripotent genes during extended period of culture in buffalo.