CAJ | 학술논문

为探索成纤维细胞作为供体细胞的潜能和Trichostatin A(TSA)处理供体细胞的适合浓度,提高克隆胚胎的发育水平.本研究分别利用100、50、25、10 ng·mL~(-1)TSA处理滩羊成纤维细胞,观察细胞形态,统计细胞活率,利用流式分选技术分析处理前后细胞的周期时相,同时通过间接免疫荧光法检测细胞乙酰化水平,并以经过10～50 ng·mL~(-1)TSA处理的成纤维细胞作为供体细胞,检测其重构胚发育水平.结果发现,当TSA的浓度达100ng·mL~(-1)时,细胞大量死亡,不同浓度TSA处理细胞24 h后,细胞周期被明显抑制在G_0/G_1期,乙酰化水平明显高于对照组,其中供体细胞经10 ng·mL~(-1) TSA处理的克隆胚胎卵裂率和囊胚率明显升高(P<0.05,(85.2±3.4)%vs(68.6±6.7)%,(35.6±5.7)%vs(10.4±8.3)%).结果表明,经10 ng·mL~(-1)TSA处理的滩羊成纤维细胞周期同步化明显,乙酰化维持较高水平,重构胚胎发育水平明显提高,适合作为成纤维细胞的处理方法和浓度.
위탐색성섬유세포작위공체세포적잠능화Trichostatin A(TSA)처리공체세포적괄합농도,제고극륭배태적발육수평.본연구분별이용100、50、25、10 ng·mL~(-1)TSA처리탄양성섬유세포,관찰세포형태,통계세포활솔,이용류식분선기술분석처리전후세포적주기시상,동시통과간접면역형광법검측세포을선화수평,병이경과10～50 ng·mL~(-1)TSA처리적성섬유세포작위공체세포,검측기중구배발육수평.결과발현,당TSA적농도체100ng·mL~(-1)시,세포대량사망,불동농도TSA처리세포24 h후,세포주기피명현억제재G_0/G_1기,을선화수평명현고우대조조,기중공체세포경10 ng·mL~(-1) TSA처리적극륭배태란렬솔화낭배솔명현승고(P<0.05,(85.2±3.4)%vs(68.6±6.7)%,(35.6±5.7)%vs(10.4±8.3)%).결과표명,경10 ng·mL~(-1)TSA처리적탄양성섬유세포주기동보화명현,을선화유지교고수평,중구배태발육수평명현제고,괄합작위성섬유세포적처리방법화농도.
In order to investigate the potential of fibroblasts as donor cells,and find out the optimal concentration of TSA used to treat fibroblasts,and further to improve the developmental level of cloned embryos,the fibroblasts were treated with 100,50,25,10 ng · mL~(-1) TSA,cell morphology and viability were investigated.Flow cytometry were adopted to analyse cell cycle.Acetylation level was detected by indirect immunofluorescence,and the developmental level of cloned embryos was evaluated.Most cells treated with 100 ng · mL~(-1) TSA were dead,but a significant increase of acetylation level was detected;the cell cycle was arrested at G_0/G_1 checkpoint with the existence of TSA;generally,there was a higher proportion of cleavages and blastulas (P <0.05,(85.2 ± 3.4)% vs( 68.6 6.7)%,(35.6 ± 5.7)% vs (10.4±8.3)%) when the cells were treated with 10 ng · mL~(-1) TSA.The results suggested that acetylation level,cell morphology and embryonic developmental level were improved and optimal for the donor cells treated with 10 ng · mL~(-1) TSA.