title: Use of peripheral blood for production of buffalo (Bubalus bubalis) embryos by handmade cloning.
abstract: Buffalo embryos were produced by handmade cloning using peripheral blood-derived lymphocytes as donor cells. Although the blastocyst rate was lower (P < 0.01) for lymphocyte- than control skin fibroblast-derived embryos (6.6 ± 0.84% vs. 31.15 ± 2.97%), the total cell number (152.6 ± 23.06 vs. 160.1 ± 13.25) and apoptotic index (6.54 ± 0.95 vs. 8.45 ± 1.32) were similar. The global level of H3K9ac was higher (P < 0.05) in lymphocyte- than that in skin-derived blastocysts; whereas in IVF blastocysts, the level was not significantly different from the two cloned groups. The level of H3K27me3 was similar among the three groups. The expression level of DNMT1, DNMT3a, HDAC1, and IGF-1R was higher (P < 0.01) in lymphocytes than that in skin fibroblasts. The expression level of CDX2 was higher (P < 0.05) than that of DNMT3a, IGF-1R, OCT4, and NANOG was lower (P < 0.05) in lymphocyte-derived than in IVF blastocysts; that of DNMT1 and HDAC1 was similar in the two groups. The expression level of all these genes, except that of NANOG, was lower (P < 0.05) in lymphocyte- than in skin fibroblast-derived blastocysts. It is concluded that, peripheral blood-derived lymphocytes can be used for producing handmade cloning embryos in bubaline buffaloes.
authors.affiliation: Animal Biotechnology Section, Central Sheep and Wool Research Institute, Avikanagar, Rajasthan, India.
authors.forename: Basanti
authors.initials: B
authors.lastname: Jyotsana
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: Amol A
authors.initials: AA
authors.lastname: Sahare
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: Anuj K
authors.initials: AK
authors.lastname: Raja
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: Karn P
authors.initials: KP
authors.lastname: Singh
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: Narendra
authors.initials: N
authors.lastname: Nala
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: S K
authors.initials: SK
authors.lastname: Singla
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: M S
authors.initials: MS
authors.lastname: Chauhan
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: R S
authors.initials: RS
authors.lastname: Manik
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India. Electronic address: prabhatpalta@yahoo.com.
authors.forename: P
authors.initials: P
authors.lastname: Palta
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002020:Buffaloes / Q000097:blood* / Q000196:embryology*; D019976:Cloning, Organism; D046149:Embryo Culture Techniques; D044127:Epigenesis, Genetic; D050437:Genes, Developmental; D012867:Skin / Q000166:cytology
keywords: Cloning; Lymphocytes; Nuclear transfer; Reprogramming; Somatic cell nuclear transfer