Universität zu Köln

Schmidt, Viola Maria, Isachenko, Evgenia, Rappl, Gunter, Rahimi, Gohar, Hanstein, Bettina, Morgenstern, Bernd, Mallmann, Peter and Isachenko, Vladimir (2018). Construction of human artificial ovary from cryopreserved ovarian tissue: Appearance of apoptosis and necrosis after enzymatic isolation of follicles. Cryobiology, 84. S. 10 - 15. SAN DIEGO: ACADEMIC PRESS INC ELSEVIER SCIENCE. ISSN 1090-2392

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Abstract

Earlier it was shown that number of retrieved follicles was significantly higher in Tumor Dissociation Enzyme (TDE)-treatment group compare to standard Liberase TM-group. The aim of our present investigations was to examine the effect of TDE on appearance of apoptosis and necrosis in follicles and stromal cells after digesting of cryopreserved ovarian cortex. Fresh and frozen ovarian cortex fragments (OCF) from 14 patients (29 +/- 6 years old), sized 20-210 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 LiberaseTM; Group 3 fresh OCF digested with TDE; Group 4 fresh OCF digested with Liberase TM. To differentiate the live, early apoptotic, late apoptotic and necrotic cells in digested ovarian cortex suspension, a flow cytometric apoptosis/necrosis assay with FITC Annexin V Apoptosis Detection Kit and with 7-AAD was performed. Most of fresh (not frozen) cells digested with TDE or Liberase TM (95 +/- 2.4% vs. 90.4 +/- 3.1%, respectively) as well as in frozen ovarian cortex digested with TDE or Liberase TM (93.1 +/- 3.4% vs. 89.7 +/- 4.4%, respectively) has located in Q3 quadrant and these cells both negative to 7-AAD and Annexin V were considered as viable. It was established that both types of enzymatic treatment applying to fresh as well as to frozen ovarian cortex resulted to high rate of viable cells (Group 1: 93.8 +/- 3.4%; Group 2: 91.8 +/- 6.0%; Group 3: 90.5 +/- 6.9%; Group 4: 87.3 +/- 2.3%) and are non significantly different (P > 0.1) between all treatment groups. The amount of early apoptotic (Group 1: 3.5 +/- 1.6%; Group 2: 4.4 +/- 1.6%; Group 3: 1.6 +/- 1.1%; Group 4: 2.4 +/- 1.5%), late apoptotic (Group 1: 2.7 +/- 2.4%; Group 2: 44.0 +/- 1.9%; Group 3: 3.1 +/- 1.1%; Group 4: 2.8 +/- 0.7% and necrotic (Group 1: 0.9% +/- 0.1%; Group 2: 2.9 +/- 0.8%; Group 3: 3.4 +/- 4.5%; Group 4: 1.1 +/- 0.6%) cells was low and was not significantly different in all treatment groups (P > 0.1). It was concluded that the use of Tumor Dissociation Enzyme, effectiveness of which is higher than Liberase TM, does not lead to increasing of apoptosis and necrosis in follicles and stromal cells after enzymatic digesting of cryopreserved ovarian cortex.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Schmidt, Viola Maria UNSPECIFIED UNSPECIFIED UNSPECIFIED Isachenko, Evgenia 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, Vladimir UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-171400
DOI:	10.1016/j.cryobiol.2018.08.011
Journal or Publication Title:	Cryobiology
Volume:	84
Page Range:	S. 10 - 15
Date:	2018
Publisher:	ACADEMIC PRESS INC ELSEVIER SCIENCE
Place of Publication:	SAN DIEGO
ISSN:	1090-2392
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CELLS; LEUKEMIA Multiple languages Biology; Physiology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/17140