Somatic cell reprogrammed by oocyte: process and barricade

Ma, Libing; He, Xiaoying; Wang, Fengmei; Cheng, Teng; Liu, XiYu

doi:10.1080/19768354.2014.927397

Cited by 3 publications

(3 citation statements)

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“…of cultured embryos) [ 23 ]. Therefore, it was considered that arresting donor cells in G0/G1 phase facilitated the development of cloned embryos derived from SCNT [ 8 ]. In previous studies of SCNT, serum starvation or full confluency was commonly used to culture nuclear donors [ 7 , 9 – 11 ], and the combination of these two methods for culturing nuclear donors was only reported in few studies [ 18 , 19 ].…”

Section: Discussionmentioning

confidence: 99%

“…In early studies, somatic cells arrested in G0/G1 phase were recommended as the ideal nuclear donors in SCNT because they facilitated coordination in the cell cycle of the somatic nuclei and the cytoplasm of oocytes [ 6 , 7 ]. Moreover, during SCNT, if the nuclear genetic material was totally removed from an oocyte, and a somatic cell in G0/G1 phase was injected (or fused) into this enucleated oocyte, after this reconstructed embryo was activated, and a certain protein synthesis inhibitor was used to prevent the exclusion of genetic material, this SCNT-derived embryo would have the correct number of chromosomes (diploid) [ 8 ]. When somatic cells are cultured in vitro , the pool of cells exists in different phases of the cell cycle.…”

Section: Introductionmentioning

confidence: 99%

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Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos

Liu

Wang

et al. 2015

PLoS ONE

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It was proposed that arresting nuclear donor cells in G0/G1 phase facilitates the development of embryos that are derived from somatic cell nuclear transfer (SCNT). Full confluency or serum starvation is commonly used to arrest in vitro cultured somatic cells in G0/G1 phase. However, it is controversial as to whether these two methods have the same efficiency in arresting somatic cells in G0/G1 phase. Moreover, it is unclear whether the cloned embryos have comparable developmental ability after somatic cells are subjected to one of these methods and then used as nuclear donors in SCNT. In the present study, in vitro cultured sheep skin fibroblasts were divided into four groups: (1) cultured to 70–80% confluency (control group), (2) cultured to full confluency, (3) starved in low serum medium for 4 d, or (4) cultured to full confluency and then further starved for 4 d. Flow cytometry was used to assay the percentage of fibroblasts in G0/G1 phase, and cell counting was used to assay the viability of the fibroblasts. Then, real-time reverse transcription PCR was used to determine the levels of expression of several cell cycle-related genes. Subsequently, the four groups of fibroblasts were separately used as nuclear donors in SCNT, and the developmental ability and the quality of the cloned embryos were compared. The results showed that the percentage of fibroblasts in G0/G1 phase, the viability of fibroblasts, and the expression levels of cell cycle-related genes was different among the four groups of fibroblasts. Moreover, the quality of the cloned embryos was comparable after these four groups of fibroblasts were separately used as nuclear donors in SCNT. However, cloned embryos derived from fibroblasts that were cultured to full confluency combined with serum starvation had the highest developmental ability. The results of the present study indicate that there are synergistic effects of full confluency and serum starvation on arresting fibroblasts in G0/G1 phase, and the short-term treatment of nuclear donor cells with these two methods could improve the efficiency of SCNT.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos

Liu

Wang

et al. 2015

PLoS ONE

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“…1.) which is considered to be appropriate for facilitating coordination in the cell cycle of the somatic nuclei, cytoplasm of oocytes and ensuring correct ploidy after nuclear transfer (Campbell et al, 1996a;Ma et al, 2014). Also, according to Campbell et al (1996a), when a cell is in quiescence, the chromatin becomes more condensed, there is a reduction in transcription and translation, and mRNAs are actively degraded.…”

Section: Figure 1 Placementioning

confidence: 99%

Technical, Biological and Molecular Aspects of Somatic Cell Nuclear Transfer – A Review

Mrowiec

Bugno‐Poniewierska

2022

Annals of Animal Science

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Since the announcement of the birth of the first cloned mammal in 1997, Dolly the Sheep, 24 animal species including laboratory, farm, and wild animals have been cloned. The technique for somatic cloning involves transfer of the donor nucleus of a somatic cell into an enucleated oocyte at the metaphase II (MII) stage for the generation of a new individual, genetically identical to the somatic cell donor. There is increasing interest in animal cloning for different purposes such as rescue of endangered animals, replication of superior farm animals, production of genetically engineered animals, creation of biomedical models, and basic research. However, the efficiency of cloning remains relatively low. High abortion, embryonic, and fetal mortality rates are frequently observed. Moreover, aberrant developmental patterns during or after birth are reported. Researchers attribute these abnormal phenotypes mainly to incomplete nuclear remodeling, resulting in incomplete reprogramming. Nevertheless, multiple factors influence the success of each step of the somatic cloning process. Various strategies have been used to improve the efficiency of nuclear transfer and most of the phenotypically normal born clones can survive, grow, and reproduce. This paper will present some technical, biological, and molecular aspects of somatic cloning, along with remarkable achievements and current improvements.

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Effects of MG132 on the in vitro development and epigenetic modification of Debao porcine somatic cell nuclear transfer embryos

KaiYuan

Dai

et al. 2017

Theriogenology

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Somatic cell reprogrammed by oocyte: process and barricade

Cited by 3 publications

References 100 publications

Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos

Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos

Technical, Biological and Molecular Aspects of Somatic Cell Nuclear Transfer – A Review

Effects of MG132 on the in vitro development and epigenetic modification of Debao porcine somatic cell nuclear transfer embryos

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