Vitamin C enhances porcine cloned embryo development and improves the derivation of embryonic stem-like cells

Fang, Xun; Tanga, Bereket Molla; Bang, Seonggyu; Seong, Gyeonghwan; Saadeldin, Islam M.; Qamar, Ahmad Yar; Shim, Joohyun; Choi, Kihwan; Lee, Sanghoon; Cho, Jongki

doi:10.1016/j.repbio.2022.100632

Cited by 12 publications

(3 citation statements)

References 63 publications

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“…The results of this study indicate that SOD1 gene expression had dramatically increased in the groups supplemented with ascorbic acid (AA) either in fresh or vitrified/thawed oocytes. This is agreement with Fang et al (2022), that reported upregulation of the antioxidant gene (SOD1) in porcine oocytes that in vitro matured in medium supplemented with ascorbic acid. According to the findings of the current study, vitrification causes lower expression in all groups whether control or AA-supplemented, than the fresh group.…”

Section: Discussionsupporting

confidence: 93%

Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes

Kandil¹,

Aboelwafa²,

Ismail³

et al. 2022

WVJ

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In vitro embryo generation, cryopreservation, and embryo transfer are examples of assisted reproductive technologies that can be used to improve camel genetic performance and fertility. The aim of this study was to investigate the impact of ascorbic acid supplementation to in vitro maturation media on the maturation rate, morphology, and gene expression of fresh and vitrified in vitro matured dromedary camel oocytes. In the current study, 810 oocytes of excellent and good quality were in vitro matured in maturation medium (TCM-199 + 10 ug/ml follicle stimulated hormone + 10% fetal calf serum + 100 IU/ml Pregnant mare serum + 50 μg/ml gentamycin) without any additives to act as a control group (C) and with 50 μg/ml ascorbic acid group (AA) and incubation in a CO2 incubator (38.5 ̊C, 5% CO2, 20% O2 and 95% humidity) for 40 hours. In vitro matured dromedary camel oocytes with the first polar body (n = 210) in C group and AA group (n = 250) were placed in basic medium (BM) and then placed in vitrification solution1 (VS1) for one minute, followed by the transfer of oocytes to VS2 (double concentration of VS1, containing 20% Ethyl Glycol (EG) and +20% Dimethyl sulfoxide) for 30 seconds. Oocytes were then loaded into sterile 0.25 ml straws and stored in liquid nitrogen for 7-10 days. The normal fresh and vitrified /thawed in vitro matured dromedary camel oocytes were kept in RNA later at a -80°C freezer for gene expression analysis. The maturation rate of dromedary camel oocytes in the in vitro matured AA group was significantly higher than that of the C group. The percentage of normally recovered vitrified/thawed oocytes was higher in the in vitro matured with ascorbic acid (VAA) than in the control (VC) group. The expression pattern of the SOD1 gene and GDF9 gene was upregulated in fresh AA and VAA groups than in the fresh C and VC groups. The profile of the SOD1 gene was more abundant in the vitrified/thawed oocytes VAA group than in the VC group. All vitrified/thawed groups, whether control or ascorbic acid supplemented, had lower levels of SOD1, GDF9, and BMP15 expression, compared to the fresh groups. In conclusion, the supplementation of the maturation medium with ascorbic acid has an increased maturation rate, and normal morphology of vitrified/ thawed oocytes which was linked with upregulation of SOD1, GDF9 genes expression.

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Section: Discussionsupporting

confidence: 93%

Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes

Kandil¹,

Aboelwafa²,

Ismail³

et al. 2022

WVJ

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“…In fact, due to limited total cell numbers and poor quality, cloned blastocysts face greater challenges in establishing ESC lines compared to in vitro fertilized or parthenogenetically activated blastocysts (40). Despite numerous attempts by researchers over the past decade, the efficiency of establishment in pigs has remained in the low single-digit percentage range (41)(42)(43). In this study, we present, for the first time, evidence that the derivation efficiency of porcine-cloned ESC lines was increased by SCF, reaching an enhancement of nearly two-fold.…”

Section: Discussionmentioning

confidence: 99%

Stem cell factor’s role in enhancing the quality of fertilized and cloned porcine embryos for improved embryonic stem cell derivation

Cai,

Hyun,

Kim

2023

Front. Vet. Sci.

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Stem cell factor (SCF), a cytokine growth factor, is expressed in various tissues of the male and female reproductive organs, including the testis, ovary, and endometrium. Its primary function involves cell survival, differentiation, and proliferation, achieved through its binding to the c-kit receptor. This study aimed to scrutinize the effects of SCF treatment during in vitro culture (IVC) on both the developmental potential and the efficiency of establishing embryonic stem cells (ESCs) from fertilized and cloned porcine embryos. The rates of cleavage and blastocyst formation exhibited no significant differences between fertilized and cloned embryos, even with the addition of SCF. However, it’s worth noting that embryos cloned with Cloud eGFP as donor cells demonstrated notably increased rates of hatched blastocysts when treated with SCF, and this increase was statistically significant (p < 0.05). Furthermore, following the complete dissection of the blastocysts, although there was no significant difference in the SCF-treated group, the area of expansion was significantly reduced (p < 0.01) in the group treated with the antagonistic blocker (ACK2) compared to both the control and SCF-treated groups. These outcomes suggest that the SCF/c-kit signaling pathway might play a pivotal role in embryo implantation. As anticipated, the efficiency of deriving ESCs was significantly higher (p < 0.01) in the group subjected to SCF treatment (12.82 ± 1.02%) compared to the control group (5.41 ± 2.25%). In conclusion, this study highlights the crucial role of SCF in enhancing the quality of porcine embryos, a vital step in obtaining high-quality ESCs.

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“…Vitamin C plays significant roles in amino acid and collagen synthesis, cholesterol metabolism, and DNA protection by acting as a free radical scavenger (12). It is also involved in many biological activities, including enzymatic reactions catalyzed by dioxygenases that use Fe 2+ and 2-oxoglutarate as co-substrates (13). Improving the culture media with sufficient vitamin C amounts leads to a more effective generation of pluripotent stem cells, somatic cells, SSCs (14), and somatic cell nuclear transfer animal embryos (3).…”

Section: Introductionmentioning

confidence: 99%

In Vitro Culture of Ovine Spermatogonial Stem Cells: Effects of Grape Seed Extracts and Vitamin C

Zolfaghari,

Zandi,

Sanjabi

et al. 2023

Gene Cell Tissue

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Background: Spermatogonial stem cells (SSCs) transmit genetic information to the next generation. For the successful transplantation of these cells, the culture medium of these cells must be optimized. Objectives: This study investigated the natural antioxidant of grape seed extract (GSE) in comparison to vitamin C in the culture medium of SSCs. Methods: A Soxhlet extractor was used to prepare ethanolic and acetonic extracts, and the Clevenger apparatus was used to prepare aqueous extracts of grape seeds. Then, their antioxidant capacities were determined using the DPPH method. The SSCs were extracted from lamb testicular tissue by a two-step digestion method, and different concentrations of GSE and vitamin C were investigated for survival, colony formation, and expression of apoptotic-related genes. Results: The results showed that the acetonic extract in the concentration of 400 µg/mL showed about 90% antioxidant properties based on the DPPH test; nevertheless, the aqueous and ethanolic extracts had only 50% of their antioxidant properties at this concentration. The acetonic extracts significantly decreased the viability of SSCs without any colony formation when used in a culture medium. The highest survival rate was obtained from the ethanolic extracts of grape seeds at the concentration of 50 µg/mL, and a significant difference was obtained with ethanolic extracts (100 to 1000 µg/mL) (P < 0.05). The viability of SSCs treated with vitamin C (50 µg/mL) was significantly higher (P < 0.05) than the control and 5 - 25 µg/mL of vitamin C groups. Conclusions: Aqueous, ethanolic, and acetonic extracts of grape seeds should not be used in the culture of SSCs. However, the use of 25 - 50 µg/mL of vitamin C is recommended in the culture of SSCs.

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Vitamin C enhances porcine cloned embryo development and improves the derivation of embryonic stem-like cells

Cited by 12 publications

References 63 publications

Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes

Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes

Stem cell factor’s role in enhancing the quality of fertilized and cloned porcine embryos for improved embryonic stem cell derivation

In Vitro Culture of Ovine Spermatogonial Stem Cells: Effects of Grape Seed Extracts and Vitamin C

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