Wang Chia-Woei, Yao Ding-Shyan, Horng Shang-Gwo, Chiu Hsiao-Chen, Chen Chun-Kai, Lee Chyi-Long, Huang Hong-Yuan, Wang Hsin-Shih, Soong Yung-Kuei, Pao Chia C
Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Lin-Kou Medical Center, 5 Fu-Hsing Street, Kwei-Shan, Tao-Yuan, Taiwan.
J Assist Reprod Genet. 2004 May;21(5):163-8. doi: 10.1023/b:jarg.0000031249.37862.3a.
The study was undertaken to test whether human telomerase reverse transcriptase (hTERT) transcripts in an individual blastomere could be used as an indicator for embryo development.
Group A consisted of day 3 normal cleaving embryos at 4- to 8-cell stage, which were surplus and not allocated for uterine transfer. Group B consisted of arrested or fragmented embryos at the same stage, which were considered to be compromised. After blastomere dissociation, RNA purification, reverse transcription, and hTERT PCR amplification, the amplified product was separated by 3% gel electrophoresis.
Eighty-six (90.5%) of the 95 intact blastomeres in group A and 78 (70.9%) of the 110 blastomeres in group B demonstrated hTERT mRNA expression. The difference was statistically significant (P < 0.05, chi-square). Eight (38.1%) of the 21 embryos in group A and 22 (62.9%) of the 35 embryos in group B had at least one blastomere that did not express hTERT mRNA under this procedure. The difference was not significant (P > 0.05, chi-square).
General hTERT mRNA transcripts can be detected in most of the individual blastomeres but cannot be used as an indicator for early embryo development. Further investigations are necessary to elucidate its clinical application.
本研究旨在测试单个卵裂球中的人端粒酶逆转录酶(hTERT)转录本是否可作为胚胎发育的指标。
A组由第3天处于4至8细胞阶段的正常分裂胚胎组成,这些胚胎为多余胚胎,未分配用于子宫移植。B组由同一阶段的停滞或碎片化胚胎组成,这些胚胎被认为存在缺陷。在卵裂球解离、RNA纯化、逆转录和hTERT PCR扩增后,扩增产物通过3%凝胶电泳分离。
A组95个完整卵裂球中有86个(90.5%)和B组110个卵裂球中有78个(70.9%)显示hTERT mRNA表达。差异具有统计学意义(P<0.05,卡方检验)。在该程序下,A组21个胚胎中有8个(38.1%)和B组35个胚胎中有22个(62.9%)至少有一个卵裂球不表达hTERT mRNA。差异无统计学意义(P>0.05,卡方检验)。
大多数单个卵裂球中可检测到一般的hTERT mRNA转录本,但不能用作早期胚胎发育的指标。需要进一步研究以阐明其临床应用。