中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2012年
5期
433-437
,共5页
李正阳%邹雨汐%孙海涛%姜晓丹
李正暘%鄒雨汐%孫海濤%薑曉丹
리정양%추우석%손해도%강효단
核移植%克隆%组蛋白去乙酰化%干细胞
覈移植%剋隆%組蛋白去乙酰化%榦細胞
핵이식%극륭%조단백거을선화%간세포
Nuclear transfer%Clone%Histone deacetylation%Stem cell
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小鼠卵母细胞为受体、卵丘细胞为供体进行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0 nmol/L(阴性对照组)、50、100、250、500 nmol/L Scriptaid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＞0.05)；250 nmol/LScriptaid组的囊胚形成率(24.2％)和囊胚细胞数(56.27±2.43)与0 nmol/L组、50 nmol/L组、100nmol/L、500 nmol/L Scriptaid组之间(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 250 nmol/L Scriptaid能有效提高小鼠SCNT克隆胚体外早期发育能力.
目的 探討抗腫瘤藥物Scriptaid對小鼠體細胞覈移植(SCNT)剋隆胚體外髮育的影響及提高移植效率的新途徑. 方法 以C57/BL6小鼠卵母細胞為受體、卵丘細胞為供體進行SCNT,構建剋隆胚,併採用數字隨機錶法將剋隆胚分為5組,分彆在含0 nmol/L(陰性對照組)、50、100、250、500 nmol/L Scriptaid的無鈣激活劑中激活6h,然後在含相應濃度Scriptaid的KSOM培養基中處理4h,最後轉移至KSOM培養基中孵育 96h.觀察、記錄各組剋隆胚的髮育情況併進行囊胚細胞計數. 結果 各組剋隆胚激活率和2-cell率之間差異無統計學意義(P＞0.05)；250 nmol/LScriptaid組的囊胚形成率(24.2％)和囊胚細胞數(56.27±2.43)與0 nmol/L組、50 nmol/L組、100nmol/L、500 nmol/L Scriptaid組之間(其囊胚形成率分彆為5.3％、6.5％、9.4％和6.9％,囊胚細胞數分彆為44.67±1.53、50.25±1.26、52.33±2.50和50.75±1.50)比較差異均有統計學意義(P＜0.05). 結論 250 nmol/L Scriptaid能有效提高小鼠SCNT剋隆胚體外早期髮育能力.
목적 탐토항종류약물Scriptaid대소서체세포핵이식(SCNT)극륭배체외발육적영향급제고이식효솔적신도경. 방법 이C57/BL6소서란모세포위수체、란구세포위공체진행SCNT,구건극륭배,병채용수자수궤표법장극륭배분위5조,분별재함0 nmol/L(음성대조조)、50、100、250、500 nmol/L Scriptaid적무개격활제중격활6h,연후재함상응농도Scriptaid적KSOM배양기중처리4h,최후전이지KSOM배양기중부육 96h.관찰、기록각조극륭배적발육정황병진행낭배세포계수. 결과 각조극륭배격활솔화2-cell솔지간차이무통계학의의(P＞0.05)；250 nmol/LScriptaid조적낭배형성솔(24.2％)화낭배세포수(56.27±2.43)여0 nmol/L조、50 nmol/L조、100nmol/L、500 nmol/L Scriptaid조지간(기낭배형성솔분별위5.3％、6.5％、9.4％화6.9％,낭배세포수분별위44.67±1.53、50.25±1.26、52.33±2.50화50.75±1.50)비교차이균유통계학의의(P＜0.05). 결론 250 nmol/L Scriptaid능유효제고소서SCNT극륭배체외조기발육능력.
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 rcconstructcd 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 of blastocysts 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＞).05).However,the form rate (24.2％) and cell numbers (56.27±2.43) of blastocysts in 250 mmol/L Scriptaid activation group were significantlyhigher 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.
