Not Available

Not AvailableBuffalo, an important Indian dairy animal, contributing significantly to milk, meat, and draught power. India is ranked first in global total milk production by producing 187 million tonnes of milk per annum, in which buffalo contributes 49% (91 million tonnes). Despite significant contributions to the country’s milk production, the number of high milk-producing buffaloes is limited. One of the major obstacles that limit the faster genetic improvement of this valued farm animal is the shortage of proven bulls to mitigate the demand of semen for breeding to produce high milk yielding buffaloes. Buffalo cloning is an asexual means of reproduction that produces genetically identical copies of a desirable buffalo without natural or artificial mating and any genome modification. Buffalo cloning is one of the assisted reproductive techniques, in which one somatic cell is fused with enucleated oocyte/oocytes to produce blastocyst stage embryos. The produced blastocysts are transferred into the foster mothers to carry cloned pregnancies and to deliver cloned calves. The application of this technology to current buffalo breeding strategies will have a substantial impact on selection intensity and subsequent genetic gain.Not Availabl

Not Available

Not AvailableThe success of in vitro fertilization (IVF), mammalian embryo culture and somatic cell nuclear transfer (SCNT) depend on stringent laboratory conditions and quality of chemicals, plastic ware, and glassware used to culture generated embryos. Therefore, all components and conditions need to be regularly monitored to avoid any kind of contamination and toxicity. Mineral oil (MO), a petroleum product, is extensively used to overlay the embryo cultures to maintain pH and osmolality of the culture medium.Not Availabl

Not Available

Not AvailableCommonly, induced pluripotent stem (iPS) cells are generated by viral transduction of four core reprogramming genes, but recent evidences suggest that slightly different combination of transcription factors improve the efficiency and quality of generated iPS cells. However, vectors like retro- and lentiviral may cause insertional mutagenesis due to its integrating ability. Hence, alternate methods with safety concerns are needed to be investigated. Therefore, the present study was undertaken to reprogram buffalo fibroblasts using non-viral piggyBac (PB) transposon mediated transfer of six transcription factors. To generate buffalo iPS cells, fibroblasts were isolated from buffalo fetus at passage 2. The cells were co-electroporated with a PB transposon having CAGGS promoter driven cassette of Oct4, Sox2, Klf4, cMyc, Nanog, and Lin28 transcription factors separated by self-cleaving 2A peptide and a helper plasmid pCMV-PB transposase. After 12-14 days post electroporation, fibroblast cells morphology was observed to change to round structures which formed loose aggregates of cells on day 18. Putative iPS cell colonies were propagated in feeder free system and characterized through expression of pluripotency markers such as alkaline phosphatase, SSEA-1, SSEA-4, SSEA-5, TRA-1-81, Oct4, Nanog and Sox2 and endogenous genes supported the stemness property of the generated cells. These cells differentiated in vitro to form embryoid bodies and were found to express three germ layers markers. In conclusion, generation of buffalo iPS cells using transposon system provides insights into viral-free iPS technology which will facilitate genetic modification of the buffalo genome and help in the production of transgenic animals using genetically modified iPS cells.Not Availabl