title: Interspecies nuclear transfer using fibroblasts from leopard, tiger, and lion ear piece collected postmortem as donor cells and rabbit oocytes as recipients.
abstract: Skin fibroblast cells were obtained from a small piece of an ear of leopard, lion, and tiger collected postmortem and attempts were made to synchronize the skin fibroblasts at G0/G1 of cell cycle using three different approaches. Efficiency of the approaches was tested following interspecies nuclear transfer with rabbit oocytes as recipient cytoplasm. Fluorescence-activated cell sorting revealed that the proportion of G0/G1 cells increased significantly (P < 0.05) when cells subjected to serum starvation, contact inhibition, and 3 mM sodium butyrate (NaBu) treatment when compared with cycling cells. However, 3 mM NaBu treatment caused alterations in cell morphology and increase in dead cells. Thus, interspecies nuclear transfer was carried out using fibroblast cells subjected to contact inhibition for 72 h, serum starvation for 48 h, and cells treated with 1.0 mM NaBu for 48 h. The fusion rates, the proportion of fused couplets that cleaved to two-cell and developed to blastocyst, were highest in all three species when the donor cells were treated with 1.0 mM NaBu for 48 h. But, the blastocyst percentage of interspecies nuclear embryos (5-6%) was significantly lower when compared with rabbit-rabbit nuclear transfer embryos (22.9%). In conclusion, fibroblast cells of leopard, lion, and tiger were successfully synchronized and used for the development of blastocysts using rabbit oocytes as recipient cytoplasm.
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, 500007, India.
authors.forename: Uma Mahesh
authors.initials: UM
authors.lastname: Yelisetti
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, 500007, India.|Transgenic Facility, EMBL Monterotondo, Adriano Buzzati-Traverso Campus, Via Ramarini 32, 00015, Monterotondo, Italy.
authors.forename: Suman
authors.initials: S
authors.lastname: Komjeti
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, 500007, India.
authors.forename: Venu Charan
authors.initials: VC
authors.lastname: Katari
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, 500007, India.
authors.forename: Shivaji
authors.initials: S
authors.lastname: Sisinthy
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, 500007, India. sambasiva@ccmb.res.in.
authors.forename: Sambasiva Rao
authors.initials: SR
authors.lastname: Brahmasani
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002453:Cell Cycle; D049109:Cell Proliferation; D047108:Embryonic Development / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology*; D005434:Flow Cytometry; D008045:Lions; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology*; D046009:Panthera; D011817:Rabbits; D013045:Species Specificity; D046010:Tigers
keywords: Cell cycle; Interspecies nuclear transfer and rabbit oocyte; Leopard; Lion; Tiger