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Updated: Jun 26, 2026
Combinational Treatment of Trichostatin A and Vitamin C Improves the Efficiency of Cloning Mice by Somatic Cell Nuclear Transfer
Published on: April 26, 2018
The developmental potential of mouse somatic cell nuclear-transferred oocytes treated with trichostatin A and
Yuta Tsuji1, Yoko Kato, Yukio Tsunoda
1Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, Japan.
Abstract:
To facilitate nuclear reprogramming, somatic cells or somatic cell nuclear-transferred (SCNT) oocytes have been treated with the histone deacetylase inhibitor trichostatin A (TSA), or the DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5-aza-dC), to relax epigenetic marks of differentiated somatic cells. TSA-treated SCNT oocytes have increased developmental potential, but the optimal treatment period is unknown. Reduced methylation levels in somatic cells have no positive effect on SCNT oocytes, but the treatment of SCNT embryos with 5-aza-dC has not been investigated. We examined the effect of TSA treatment duration on the developmental potential of mouse SCNT oocytes and the effect of 5-aza-dC treatment on their in vitro and in vivo developmental potential. To determine the effects of TSA treatment duration, nuclear-transferred (NT) oocytes were cultured for 0 to 26 h with 100 nM TSA. SCNT oocytes treated with TSA for 8 to 12 h had the higher rate of development to blastocysts and full-term fetuses were obtained after treatment for 8 to 12 h. When oocytes were treated for 14 h and 26 h, blastocyst rates were significantly decreased and fetuses were not obtained. To examine the effect of 5-aza-dC, 2-cell stage SCNT embryos were cultured with 10 or 100 nM 5-aza-dC for 48 h to the morula stage and transferred. The potential of embryos treated with 5-aza-dC to develop into blastocysts was decreased and no fetuses were obtained after transfer. The findings demonstrated that long-term TSA treatment of SCNT mouse oocytes and treatment with 5-aza-dC inhibit the potential to develop into blastocysts and to fetuses after transfer.
Insights
Optimal treatment with trichostatin A (TSA) for somatic cell nuclear transfer (SCNT) oocytes is 8–12 hours, enhancing developmental potential. Longer TSA or 5-aza-2'-deoxycytidine (5-aza-dC) treatment inhibits SCNT embryo development.
Area of Science:
- Reproductive Biology
- Epigenetics
- Developmental Biology
Background:
- Somatic cell nuclear transfer (SCNT) requires epigenetic reprogramming of differentiated cells.
- Histone deacetylase inhibitors like trichostatin A (TSA) and DNA methyltransferase inhibitors like 5-aza-2 -deoxycytidine (5-aza-dC) are used to relax epigenetic marks.
- The optimal duration of TSA treatment and the effect of 5-aza-dC on SCNT oocytes/embryos remain unclear.
Purpose of the Study:
- To determine the optimal treatment period of TSA for mouse SCNT oocytes to maximize developmental potential.
- To investigate the effect of 5-aza-dC treatment on the in vitro and in vivo developmental potential of mouse SCNT embryos.
Main Methods:
- SCNT oocytes were cultured with 100 nM TSA for varying durations (0–26 hours).
- SCNT embryos at the 2-cell stage were cultured with 10 or 100 nM 5-aza-dC for 48 hours to the morula stage.
- Developmental potential was assessed by blastocyst formation rates and full-term fetus development after embryo transfer.
Main Results:
- TSA treatment for 8–12 hours resulted in the highest blastocyst rates and full-term fetuses.
- Extended TSA treatment (14–26 hours) significantly decreased blastocyst rates and prevented fetus development.
- 5-aza-dC treatment of SCNT embryos decreased blastocyst formation and yielded no fetuses.
Conclusions:
- An 8–12 hour treatment period with TSA is optimal for enhancing the developmental potential of mouse SCNT oocytes.
- Prolonged TSA treatment and 5-aza-dC treatment negatively impact the developmental capacity of SCNT embryos.
- These findings highlight the critical importance of treatment duration and specific epigenetic modifiers in SCNT efficiency.
