Lee Min Jin, Kim Soo Hyun, Shim Sung Han, Jang Hee Yeon, Park Hee Jin, Cha Dong Hyun
Department of Obstetrics and Gynecology, CHA Ilsan Medical Center, CHA University, Gyenoggi-do KS009, Korea.
Department of Obstetrics and Gynecology, CHA Gangnam Medical Center, CHA University, Seoul KS013, Korea.
Diagnostics (Basel). 2020 May 14;10(5):300. doi: 10.3390/diagnostics10050300.
Extravillous trophoblast cells (EVTs) secreted by the uterine cavity may help overcome limitations associated with prenatal testing currently in use. EVTs are isolated using a routine safe liquid-based Pap test (called ThinPrep); however, the ThinPrep solution contains alcohol that hinders the isolation of intact EVTs. We compared the trophoblastic cell isolation efficiency of two different methods of fixation: Thinprep (pre-fixation method) and formalin (post-fixation method). We analyzed EVTs from 20 pregnant women (5-20 weeks of gestation) who underwent invasive prenatal testing. The percentages of placental β-human chorionic gonadotropin (β-hCG)-expressing cells were calculated. The presence of XY chromosomes were used to confirm pure trophoblast cells by fluorescence in situ hybridization. The β-hCG-positive cells obtained from pre- and post-fixation were 66.4 ± 13.3% and 83.2 ± 8.1% ( = 0.003), respectively, and fluorescence-positive cells were 11.1 ± 2.1% and 23.8 ± 4.8%, respectively ( = 0.001). Post-fixation was found to be more efficient in isolating non-trophoblast cells than pre-fixation. For the successful clinical application of trophoblast retrieval and isolation from the cervix in prenatal genetic testing, each step should be optimized for consistent and reliable results.
子宫腔分泌的绒毛外滋养层细胞（EVTs）可能有助于克服目前使用的产前检测相关的局限性。EVTs是使用常规安全的液基巴氏试验（称为ThinPrep）分离的；然而，ThinPrep溶液中含有酒精，这会阻碍完整EVTs的分离。我们比较了两种不同固定方法的滋养层细胞分离效率：Thinprep（预固定法）和福尔马林（后固定法）。我们分析了20名接受侵入性产前检测的孕妇（妊娠5 - 20周）的EVTs。计算了胎盘β - 人绒毛膜促性腺激素（β - hCG）表达细胞的百分比。通过荧光原位杂交使用XY染色体的存在来确认纯滋养层细胞。预固定和后固定获得的β - hCG阳性细胞分别为66.4±13.3%和83.2±8.1%（P = 0.003），荧光阳性细胞分别为11.1±2.1%和23.8±4.8%（P = 0.001）。发现后固定在分离非滋养层细胞方面比预固定更有效。为了在产前基因检测中从宫颈成功临床应用滋养层细胞的检索和分离，每个步骤都应进行优化以获得一致且可靠的结果。
