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Chromosomal analysis of meiotic stages of human oocytes matured in vitro: benefits of protease treatment before fixation.

Abstract：

OBJECTIVE:To modify Tarkowski's air-dry technique for mouse oocytes to develop a rapid, consistent procedure for human oocytes that enables accurate scoring of meiotic stage. DESIGN, SETTING, AND PATIENTS:Meiotically immature human oocytes, obtained after oophorectomy, were cultured for various periods and then subjected to Tarkowski's air-dry procedure (n = 104) or to our modified procedure (n = 175) that used a brief exposure to protease (20 to 40 seconds) before fixation. MAIN OUTCOME MEASURES:Air-dried oocytes were assessed for readability and for whether they contained overspread or overlapping chromosomes. In addition, discrete meiotic stages in human oocytes were identified. RESULTS:Our protease procedure significantly increased readability of air-dried oocytes (96% versus 79% readable for protease versus Tarkowski, respectively; P less than 0.001) by significantly reducing the number of preparations with either overscattered (0.7% versus 3.4% for protease versus Tarkowski, respectively, P less than 0.05) or overlapping (1.3% versus 18% for protease versus Tarkowski, respectively, P less than 0.001) chromosomes. CONCLUSIONS:Protease exposure of oocytes, combined with a modification of Tarkowski's procedure, resulted in high quality air-dries of human oocytes. This rapid and reliable procedure should have clinical application in in vitro fertilization programs for meiotic assessment of oocytes failing to fertilize.

journal_name

Fertil Steril

journal_title

Fertility and sterility

authors

Racowsky C,Kaufman ML,Dermer RA,Homa ST,Gunnala S

doi

10.1016/s0015-0282(16)55021-3

keywords:

subject

Has Abstract

pub_date

1992-05-01 00:00:00

pages

1026-33

issue

5

eissn

0015-0282

issn

1556-5653

pii

S0015-0282(16)55021-3

journal_volume

57

pub_type

杂志文章

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