Use of peripheral blood for production of buffalo (Bubalus bubalis) embryos by handmade cloning

Jyotsana, Basanti; Sahare, Amol Ashok; Raja, A; Singh, Kiran; Nala, Narendra; Singla, S. K.; Chauhan, M. S.; Manik, R. S.; Palta, P.

doi:10.1016/j.theriogenology.2016.04.073

Cited by 12 publications

(4 citation statements)

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“…In general, as observed in other mammals (Guan et al, 2010;Kwong et al, 2014), establishment of an adequate cell line is a prerequisite step for the success of cloning and producing induced pluripotent cells . For these techniques, fibroblasts and epithelial cells derived from the skin have been widely used (Jyotsana et al, 2016;Siengdee et al, 2018). Initially, epithelial and fibroblast cells were grown simultaneously; nevertheless, fibroblasts can more easily adhere as well as detach by trypsinization as compared to the epithelial cells (Bai et al, 2012;Saadeldin et al, 2019;Siengdee et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines

Borges

Lira

Nascimento

et al. 2020

PeerJ

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Background Biobanking of cell lines is a promising tool of support for wildlife conservation. In particular, the ability to preserve fibroblast cell lines derived from collared peccaries is of significance as these wild mammals are unique to the Americas and play a large role in maintaining the ecosystem. We identified collared peccary fibroblasts by immunofluorescence and evaluated their morphology, growth and adherence capacity. Further, we monitored the viability and metabolic activity of the fibroblasts to determine the effects of passage number and cryopreservation on establishment of cell lines. Methods Skin biopsies were collected from the peripheral ear region from five adult animals in captivity. Initially, cells were isolated from fragments and cultured in the Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum and 2% antibiotic–antimycotic solution under a controlled atmosphere (38.5 °C, 5% CO2). We evaluated the maintenance of primary cells for morphology, adherence capacity of explants, explants in subconfluence, cell growth and absence of contamination. Moreover, we identified the fibroblast cells by immunofluorescence. Additionally, to evaluate the influence of the number of passages (first, third and tenth passage) and cryopreservation on establishment of cell lines, fibroblasts were analysed for the viability, metabolic activity, population doubling time (PDT), levels of reactive oxygen species (ROS), and mitochondrial membrane potential (ΔΨm). Results All explants (20/20) adhered to the dish in 2.4 days ± 0.5 with growth around the explants in 4.6 days ± 0.7, and subconfluence was observed within 7.8 days ± 1.0. Moreover, by morphology and immunocytochemistry analyses, cells were identified as fibroblasts which presented oval nuclei, a fusiform shape and positive vimentin staining. No contamination was observed after culture without antibiotics and antifungals for 30 days. While there was no difference observed for cell viability after the passages (first vs. third: P = 0.98; first vs. tenth: P = 0.76; third vs. tenth: P = 0.85), metabolic activity was found to be reduced in the tenth passage (23.2 ± 12.1%) when compared to that in the first and third passage (100.0 ± 24.4%, P = 0.006). Moreover, the cryopreservation did not influence the viability (P = 0.11), metabolic activity (P = 0.77), or PDT (P = 0.11). Nevertheless, a greater ΔΨm (P = 0.0001) was observed for the cryopreserved cells (2.12 ± 0.14) when compared to that in the non-cryopreserved cells (1.00 ± 0.05). Additionally, the cryopreserved cells showed greater levels of intracellular ROS after thawing (1.69 ± 0.38 vs. 1.00 ± 0.22, P = 0.04). Conclusions This study is the first report on isolation, characterization and cryopreservation of fibroblasts from collared peccaries. We showed that adherent cultures were efficient for obtaining fibroblasts, which can be used as donor cells for nuclei for species cloning and other applications.

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Section: Introductionmentioning

confidence: 99%

Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines

Borges

Lira

Nascimento

et al. 2020

PeerJ

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“…Epigenetic modifications such as DNA methylation and histone modifications play an important role in embryonic development (Niemann, 2016 ; Sproul et al, 2005 ). Aberrant epigenetic modifications such as DNA methylation and histone acetylation, and also abnormal gene expression patterns for example insulin‐like growth factors (IGF‐1 and IGF‐2) have been found in cloned buffalo embryos when compared with those of IVF embryos (Jyotsana et al, 2016 ; Luo et al, 2013 ; Mohapatra et al, 2015 ; Pandey et al, 2009 ; Saini et al, 2016 , 2017 ; Srirattana et al, 2014 ; Sun et al, 2015 ; Suteevun, Parnpai, et al, 2006 ; Suteevun, Smith, et al, 2006 ).…”

Section: Scntmentioning

confidence: 99%

Current status of assisted reproductive technologies in buffaloes

Srirattana

Hufana-Durán

Atabay

et al. 2022

Animal Science Journal

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Buffaloes are raised by small farm holders primarily as source of draft power owing to its resistance to hot climate, disease, and stress conditions. Over the years, transformation of these animals from draft to dairy was deliberately carried out through genetic improvement program leading to the development of buffalo-based enterprises. Buffalo production is now getting more attention and interest from buffalo raisers due to its socioeconomic impact as well as its contribution to propelling the livestock industry in many developing countries. Reproduction of buffaloes, however, is confronted with huge challenge and concern as being generally less efficient to reproduce compared with cattle due to both intrinsic and extrinsic factors such as poor estrus manifestation, silent heat, marked seasonal infertility, postpartum anestrus, long calving interval, delayed puberty, inherently low number of primordial follicles in their ovaries, high incidence of atresia, and apoptosis. Assisted reproductive technologies (ARTs) are major interventions for the efficient utilization of follicle reserve in buffaloes. The present review focuses on estrus and ovulation synchronization for fixed time artificial insemination, in vitro embryo production, intracytoplasmic sperm injection, cryopreservation of oocytes and embryos, somatic cell nuclear transfer, the factors affecting utilization in various ARTs, and future perspectives in buffaloes.

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“…On the other hand, SCNT faces major problems of low efficiency and high mortality rates of fetuses and embryos (Kishigami et al, 2008). There are a number of factors that influence the efficiency of SCNT which includes the donor cell type (Oback and Wells, 2007;Salamone, 2006), enucleation protocol (Moro et al, 2010), electrofusion and activation steps (Canel et al, 2010;Vichera et al, 2009 (George et al, 2011;Arat et al, 2002;Huang et al, 2010;Jyotsana et al, 2015;Madheshiya et al,2015). But till now, it is not clear which cell type is best suited for cloning (Kato et al, 1998;Goto et al, 2013;Tani et al, 2001).…”

Section: Somatic Cell Nuclear Transfer (Scnt)mentioning

confidence: 99%

Transgenesis in farm animals-A review

Mehta

Sharma

Kaushik

2017

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For a number of decades, attempts have been made to successfully produce transgenic animals which have numerous applications in the biotechnology industry with the foremost emphasis on production of monoclonal antibodies and recombinant proteins of human welfare. Different techniques are adopted in order to produce transgenic farm animals which could be further used as bioreactors. The most common traditional transgenesis technique employed is Somatic Cell Nuclear Transfer (SCNT) using genetically modified somatic cells or stem cells as nuclear donors. This review article summarizes the merits and demerits of the techniques currently used to produce transgenic livestock with major emphasis on somatic cell nuclear transfer. In the end, a brief discussion is done about the novel methods adopted to produce transgenic animals like Zinc Finger Nucleases (ZFN), Transcription Activator-like Effector Nuclease (TALEN) and Clustered regularly interspaced short palindromic repeats (CRISPR). It is expected that the new techniques developed would overcome the problems faced with existing traditional transgenesis methods.

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Use of peripheral blood for production of buffalo (Bubalus bubalis) embryos by handmade cloning

Cited by 12 publications

References 30 publications

Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines

Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines

Current status of assisted reproductive technologies in buffaloes

Transgenesis in farm animals-A review

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