Universität zu Köln

Schmidt, Viola Maria, Isachenko, Vladimir, Rappl, Gunter, Rahimi, Gohar, Hanstein, Bettina, Morgenstern, Bernd, Mallmann, Peter and Isachenko, Evgenia (2018). Comparison of the enzymatic efficiency of Liberase TM and tumor dissociation enzyme: effect on the viability of cells digested from fresh and cryopreserved human ovarian cortex. Reprod. Biol. Endocrinol., 16. LONDON: BIOMED CENTRAL LTD. ISSN 1477-7827

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Abstract

Background: The aim of this study was to examine the effectiveness of Tumor Dissociation Enzyme (TDE) on the viability of follicles after digestion of fresh and cryopreserved ovarian cortex fragments (OCFs). Methods: Fresh and thawed OCF from 14 patients (29 +/- 6 years), sized 20 to 210 mm(3) were randomly distributed into four treatment groups and digested with 16% TDE or 0.05 mg/ml Liberase TM: Group 1, frozen OCF digested with TDE; Group 2, frozen OCF digested with Liberase TM; Group 3, fresh OCF digested with TDE; and Group 4, fresh OCF digested with Liberase TM. Evaluation of follicle viability was performed under light microscope after staining with Neutral red. For visualization of viable and dead cells under a confocal laser scanning microscope, the follicles were stained with Calcein AM and ethidium homodimer-1. Results: The results showed that the number of retrieved follicles was significantly higher (990 vs 487; P < 0.01) in the TDE-treatment group compared to the Liberase TM-group. The presence of intense neutral red stained follicles was significantly higher in Group 1 and Group 3 compared to Group 2 and Group 4 (70.3% +/- +/-6.22 vs 53,1% +/- 2.03 and 94.2% +/- 6.6 vs 79.1% +/- 2.1; P < 0.01). The percentage of Calcein AM stained follicles of class V1 was significantly higher in Group 1 and Group 3 compared to Group 2 and Group 4 (95.97% +/- 7.8 vs 87.87% +/- 2.4; 97.1% +/- 6.8 vs 91.3% +/- 2.3; P < 0.01). Conclusion: The enzymatic digestion of ovarian cortex with TDE provides recovery of a higher number of healthy preantral follicles in contrast to earlier described Liberase TM procedure.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Schmidt, Viola Maria UNSPECIFIED UNSPECIFIED UNSPECIFIED Isachenko, Vladimir UNSPECIFIED UNSPECIFIED UNSPECIFIED Rappl, Gunter UNSPECIFIED UNSPECIFIED UNSPECIFIED Rahimi, Gohar UNSPECIFIED UNSPECIFIED UNSPECIFIED Hanstein, Bettina UNSPECIFIED UNSPECIFIED UNSPECIFIED Morgenstern, Bernd UNSPECIFIED UNSPECIFIED UNSPECIFIED Mallmann, Peter UNSPECIFIED UNSPECIFIED UNSPECIFIED Isachenko, Evgenia UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-183112
DOI:	10.1186/s12958-018-0374-6
Journal or Publication Title:	Reprod. Biol. Endocrinol.
Volume:	16
Date:	2018
Publisher:	BIOMED CENTRAL LTD
Place of Publication:	LONDON
ISSN:	1477-7827
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PRE-ANTRAL FOLLICLES; HUMAN PREANTRAL FOLLICLES; LONG-TERM CULTURE; IN-VITRO; PRIMORDIAL FOLLICLES; FOLLICULAR-GROWTH; NEUTRAL RED; QUANTITATIVE-ANALYSIS; OOCYTE MATURATION; CANCER-PATIENTS Multiple languages Endocrinology & Metabolism; Reproductive Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18311