Scriptaid对小鼠体细胞核移植克隆胚早期发育能力的提高作用

Scriptaid improves early development potential of mouse somatic cell nuclear transfer embryos

目的探讨抗肿瘤药物Scriptaid对小鼠体细胞核移植（SCNT）克隆胚体外发育的影响及提高移植效率的新途径。方法以C57／BL6小鼠卯母细胞为受体、卯丘细胞为供体进行SCNT，构建克隆胚，并采用数字随机表法将克隆胚分为5组，分别在含0nmol／L（阴性对照组1、50、100、250、500nmol／LScriptaid的无钙激活剂中激活6h．然后在含相应浓度Scriptaid的KSOM培养基中处理4h，最后转移至KSOM培养基中孵育96h。观察、记录各组克隆胚的发育情况并进行囊胚细胞计数。结果各组克隆胚激活率和2．cell率之间差异无统计学意义（P〉O．05）；250nmol／LScript,aid组的囊胚形成率（24．2％）和囊胚细胞数（56．27±2．43）与0 nmol／L组、50nmol／L组、100nmol／L、500nmol／LScriptaid组之间f其囊胚形成率分别为5．3％、6．5％、9．4％和6．9％．囊胚细胞数分别为44.67±1.53, 50.25±1.26, 52.33± 2.50 和 50.75±1.50比较差异均有统计学意义（P〈0．05）。结论250nmol／LScrimaid能有效提高小鼠SCNT克隆胚体外早期发育能力。

Objective To examine the effect of Scriptaid, a histone deacetylase inhibitor and a kind of anti-cancer drug, on development of mouse somatic cell nuclear transfer （SCNT） embryos in vitro and explore a new strategy to improve the efficiency of SCNT. Methods SCNT was carried out by pizeo-activated micromanipulator in C57/BL6 mouse, from which the oocytes were chsoen as recipients and the cumulus cells as donors. The reconstructed embryos were randomly divided into 5 groups and activated in calcium-free activators with 0 mmol/L （negative control group）, and 50, 100, 250 and 500 mmol/L Scriptaid for 6 h, respectively; and then, they were transferred into KSOM medium with corresponding concentrations of Scriptaid for 4 h. The cloned embryos were finally cultured in KSOM medium for 96 h. The development （form rate） of cloned embryos and the count ofblastocysts cells were recorded. Results No significant differences on the activated rate of the reconstructed embryos and the 2-cell cleavage rate were noted between each 2 groups （P〉0.05）. However, the form rate （24.2%） and cell numbers （56.27±2.43） of blastocysts in 250 mmol/L Scriptaid activation group were significantly higher as compared with those in the negative control group, and 50, 100 and 500 mmol/L Scriptaid activation groups （form rates： 5.3%, 6.5%, 9.4% and 6.9%; cell numbers： 44.67±1.53, 50.25±1.26, 52.33± 2.50 and 50.75±1.50, respectively, P〈0.05）. Conclusion The early development potential of mouse SCNT embryos in vitro can be dramatically improved by 250mmol/L Scriptaid.