采用原位末端标记法检测人植入前胚胎中的凋亡征象

Detection of apoptosis in human preimplantation embryos by TdT-mediated nick end labeling

目的建立原位末端标记法(TdT-mediated nick end labeling TUNEL)检测人植入前胚胎凋亡征象的方法,阐明凋亡与人类植入前胚胎发育的关系。方法取试管婴儿助孕技术后异常受精的三原核(3PN)胚胎,于2-10细胞期进行固定,采用TUNEL法检测3PN胚胎中的凋亡征象。结果在发育正常3PN胚胎中,未检测到凋亡征象;在20%有碎片的3PN胚胎中检测到TUNEL阳性信号。结论采用TUNEL法可以有效检测人植入前胚胎中的凋亡征象,植入前胚胎中的细胞碎片可能与凋亡有关。

Objective： To establish the detection method of apoptosis in human preimplantation embryos by TdT- mediated nick end labeling （TUNEL） and elucidate the relationship of apoptosis to development of human preimplantation embryos. Methods： The three pronuclear （3PN） human preimplantation embryos were fixed at 2 -10 cell stage. TdT- mediated nick end labeling and laser eonfocal scanning microscopy were used to detect apoptosis signs in embryos. Results： No positive TUNEL signals were found in normal 3PN embryos. The TUNEL assay was positive in 19% of fragmented embryos. Conclusion： TUNEL is an effective method to detect apoptosis in human preimplantation embryos. Fragments in embryos are possibly related to apoptosis.