Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos

Huang, Yongye; Tang, Xiaochun; Xie, Wanhua; Zhou, Yan; Dong, Liang; Zhou, Yang; Zhu, Junyan; Yuan, Ting; Lai, Liangxue; Pang, Daxin; Ouyang, Hongsheng

doi:10.1016/j.bbrc.2011.06.160

Cited by 76 publications

(74 citation statements)

References 22 publications

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“…We found that 30 mg/ml of vitamin C had a better effect on the proliferation of ADSCs than 60 and 90 mg/ml of vitamin C. The ADSCs treated with 30 mg/ml of vitamin C have demonstrated the best cell proliferation activity, which was similar to previous reports about the concentration-dependent effect of vitamin C on cell proliferation. [18][19][20] In addition, our findings also indicated that cell cycle progression was promoted, and the decreased percentage of G 1 phase was obviously observed after incubation with 30 mg/ml of vitamin C for 24 and 48 h, and meanwhile, the increased percentage of the cells at S and G 2 /M phase was also observed. To further investigate the mechanism by which vitamin C increased cell proliferation and promoted cell cycle progression, we detected the cell cycle-associated proteins including cyclin E1, p53, p21, and CDK2.…”

Section: Discussionsupporting

confidence: 61%

Vitamin C promotes the proliferation of human adipose-derived stem cells via p53-p21 pathway

Zhang

et al. 2016

Organogenesis

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ABSTRACT. Although adipose-derived stem cells (ADSCs) have demonstrated a promising potential for the applications of cell-based therapy and regenerative medicine, excessive reactive oxygen species (ROS) are harmful to ADSCs cell survival and proliferation. Vitamin C is an important antioxidant, and is often added into culture media as an essential micronutrient. However, its roles on the proliferation of human ADSCs have not been studied. Therefore, in this study, human ADSCs were isolated, and detected by flow cytometry for the analysis of their cell surface antigens. Cell proliferation and cell cycle progression were measured with cell counting kit-8 assay and flow cytometry, respectively. Western blotting was used to detect the expression levels of cyclin E1, p53, p21, and CDK2 proteins. The effect of vitamin C pretreatment on the production of hydrogen peroxide (H 2 O 2 )-mediated ROS in the ADSCs was evaluated by flow cytometry. Our results indicated that vitamin C treatment significantly increased cell proliferation, and changed the cell cycle distribution of ADSCs by decreasing the percentage of G 1 phase, and concurrently increased the percentage of S and G 2 /M phase. Western blot analysis indicated that vitamin C treatment up-regulated the expression levels of cyclin E1 and CDK2, but down-regulated p53 and p21 proteins expression, which contributed to cell proliferation and cell cycle progression. Vitamin C pretreatment significantly reduced the production of H 2 O 2 -induced ROS in the ADSCs. These findings suggest that vitamin C can promote the proliferation and cell cycle progression in the ADSCs possibly through regulation of p53-p21 signal pathway.

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Section: Discussionsupporting

confidence: 61%

Vitamin C promotes the proliferation of human adipose-derived stem cells via p53-p21 pathway

Zhang

et al. 2016

Organogenesis

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“…The active form of the vitamin E homologues is α-tocopherol which protects cell membranes from oxidation by reacting with ROS and lipid radicals produced in the lipid peroxidation. Supplementation of vitamins to culture medium improves embryo development in mouse [117], pig [38, 118,119] and cow [120].…”

Section: Development Of Embryosmentioning

confidence: 99%

Oxidative Stress and Redox Regulation on <i>In Vitro</i> Development of Mammalian Embryos

Takahashi

2012

Journal of Reproduction and Development

204

184

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Abstract. Many factors affect development of mammalian preimplantation embryos in vitro. It is well known that in vitro development of bovine embryos is highly affected by culture condition including energy source, growth factors, pH or gas environment. Many efforts have been made towards the suitable environments which can successfully support embryo development in vitro. For a rapid growth and differentiation, embryo requires energy by utilizing ATP, NADPH with oxygen molecules. These energy substrates are produced from the electron transport chain in the mitochondria. In addition to energy production, reactive oxygen species (ROS) are also generated as by-product of such energy production system. ROS production is sensitively controlled by the balance of oxidizing and reducing status and affected by several antioxidant enzymes such as superoxide dismutase (SOD), Catalase, glutathione peroxidase (GPx) or low molecular weight thiols such as glutathione (GSH). Imbalance of oxidation and reduction causes production of excess ROS, which causes the developmental arrest, physical DNA damage, apoptosis induction or lipid peroxidation. Environmental oxygen condition during embryo culture also highly affects embryo development as well as intracellular redox balance. Several studies have revealed that regulation of intra-and extra-cellular reducing environment by reducing excess ROS by using antioxidants, reducing oxygen concentration are effective for improving embryo development. Also, recent studies have demonstrated the difference in gene expression affected by oxidative stress. This review briefly summarizes the effects of ROS and the role of redox balance on preimplantation embryos for improving the efficiency of in vitro production of mammalian embryos. Key words: Embryo development, Glutathione, Intracellular redox state, Reactive oxygen species (J. Reprod. Dev. 58: 1-9, 2012) H andling of oocytes and embryos in vitro is an effective technique for obtaining offspring by embryo transfer, investigating the mechanisms of early embryonic development and differentiation, gene functions by transgenic approach and research of somatic cell clones in mammals. To improve the pregnancy rates in in vitro produced (IVP) embryos, it is important to evaluate and improve the quality of embryos suitable for transfer. Therefore, increasing the number of good quality embryos as well as selection of superior quality embryos is one of the most practical and clinically useful approaches in embryo transfer of mammalian preimplantation embryos. It is well known that in vitro development of bovine embryos is highly affected by culture condition. Therefore, much effort has been made towards the effective culture methods that support embryo development in vitro. The detrimental effect of oxidative stress caused by a high oxygen concentration under culture condition on embryo development has been studied with the comparison of oxygen concentration in in vivo condition.It was found that embryo development is promoted under a low O 2 concent...

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“…For producing cloned embryos, four types of anti-CSFV shRNA-positive clonal cells, scrambled shRNA clonal cells, and normal control cells were transferred into enucleated oocyte recipient cells at the metaphase II stage. Then, the SCNT embryos were cultured in PZM3 for 148 h. The development of SCNT embryos was evaluated by measuring the cleavage and blastocyst formation rates (Huang et al, 2011). In our experiment, the rate of blastocyst formation did not show significant difference between the experimental and control groups ( Table 1), suggesting that adverse effects caused by shRNA may occur in later developmental stages.…”

Section: Blastocyst Formation Rate For Donor Cells Stably Transfectedmentioning

confidence: 69%

Early lethality of shRNA-transgenic pigs due to saturation of microRNA pathways

Dai

Zhao

et al. 2014

J. Zhejiang Univ. Sci. B

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RNA interference (RNAi) is considered as a potential modality for clinical treatment and anti-virus animal breeding. Here, we investigate the feasibility of inhibiting classical swine fever virus (CSFV) replication by short hairpin RNA (shRNA) in vitro and in vivo. We generate four different shRNA-positive clonal cells and two types of shRNA-transgenic pigs. CSFV could be effectively inhibited in shRNA-positive clonal cells and tail tip fibroblasts of shRNA-transgenic pigs. Unexpectedly, an early lethality due to shRNA is observed in these shRNA-transgenic pigs. With further research on shRNA-positive clonal cells and transgenic pigs, we report a great induction of interferon (IFN)-responsive genes in shRNA-positive clonal cells, altered levels of endogenous microRNAs (miRNA), and their processing enzymes in shRNA-positive cells. What is more, abnormal expressions of miRNAs and their processing enzymes are also observed in the livers of shRNA-transgenic pigs, indicating saturation of miRNA/shRNA pathways induced by shRNA. In addition, we investigate the effects of shRNAs on the development of somatic cell nuclear transfer (SCNT) embryos. These results show that shRNA causes adverse effects in vitro and in vivo and shRNAinduced disruption of the endogenous miRNA pathway may lead to the early lethality of shRNA-transgenic pigs. We firstly report abnormalities of the miRNA pathway in shRNA-transgenic animals, which may explain the early lethality of shRNA-transgenic pigs and has important implications for shRNA-transgenic animal preparation.

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Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos

Cited by 76 publications

References 22 publications

Vitamin C promotes the proliferation of human adipose-derived stem cells via p53-p21 pathway

Vitamin C promotes the proliferation of human adipose-derived stem cells via p53-p21 pathway

Oxidative Stress and Redox Regulation on <i>In Vitro</i> Development of Mammalian Embryos

Early lethality of shRNA-transgenic pigs due to saturation of microRNA pathways

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