Follicles of various maturation stages react differently to enzymatic isolation: a comparison of different isolation protocols

REPRODUCTIVE BIOMEDICINE ONLINE

卷:30

Article

Lierman, Sylvie¹  Tilleman, Kelly¹  Cornelissen, Maria²  De Vos, Winnok H.^3,4  Weyers, Steven⁵  T'Sjoen, Guy⁶  Cuvelier, Claude A.⁷  De Sutter, Petra¹ 

[1] Ghent Univ Hosp, Dept Reprod Med, B-9000 Ghent, Belgium

[2] Univ Ghent, Fac Med & Hlth Sci, Dept Basic Med Sci, B-9000 Ghent, Belgium

[3] Univ Antwerp, Fac Biochem Pharmaceut & Vet Sci, Dept Vet Sci, B-2020 Antwerp, Belgium

[4] Univ Ghent, Fac Biosci Engn, Dept Mol Biotechnol, B-9000 Ghent, Belgium

[5] Ghent Univ Hosp, Dept Gynaecol, B-9000 Ghent, Belgium

[6] Ghent Univ Hosp, Dept Endocrinol, B-9000 Ghent, Belgium

[7] Ghent Univ Hosp, Dept Pathol, B-9000 Ghent, Belgium

关键词: apoptosis;    enzyme;    follicle isolation;    human;    ovarian tissue;    vitality;   

DOI  :  10.1016/j.rbmo.2014.10.009

来源: Elsevier

【 摘 要 】

Isolation of human follicles is based on digestion of the tissue by combinations of enzymes. Follicle vitality and morphology are often based on the analysis of pooled follicles of different maturation stages. Information is therefore lacking on the effect of the isolation protocol to individual follicles of different maturation stages. A study was conducted using five protocols combining different enzymes and varying concentrations. Isolated follicles were classified according to their maturation stages, counted and characterized for vitality, morphology, early apoptosis and organization of transzonal projections. No statistical differences were found between the protocols when outcome parameters were analysed on a pool of follicles regardless of their maturation status. Differences were observed in quality when the follicles were analysed separately according to their maturation status. Combining morphologic characteristics and vitality, both Liberase DH and Liberase TM combined with collagenase IV were better at isolating high-quality primordial follicles, compared with collagenase IV. No statistical difference between the isolation protocols was found for primary follicles. If only high-quality isolated secondary follicles are needed, collagenase IV is found to be most advantageous. Follicles of different maturation stages react differently when enzymatic isolation protocols are compared. (C) 2014 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

【 授权许可】

Free