Comparison of sucrose and trehalose media modification as an update of oocyte vitrification: A study of apoptotic level

Silvia Werdhy Lestari, Nurin N. Fitriyah, Mulyoto Pangestu, Gita Pratama, Ria Margiana

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

Number of women who are not being able to have offspring in their reproductive life is increasing which might be influenced by several factors. As a consequence, oocyte cryopreservation could be an ensuring solution for women fertility preservation. A good vitrification could be conducted by combining an appropriate of type and concentration of cryoprotectants. One of the marks of successful vitrification is the vitrified oocytes could avoid apoptosis. This study aimed to evaluate the modification of cryoprotectant media as un update of oocyte vitrification as follow: the combination and the concentration of cryoprotectant media of oocytes vitrification, based on their effects on the apoptosis or DNA damage of oocytes. A total of 84 MII stage oocytes from adult female Deutschland, Denken and Yoken (DDY) mice (7-8 weeks old) were used in this study. Vitrification procedure was performed by using VS1 contained 15% EG, 15% DMSO, 0.5 mol/l sucrose (Merck, Darmstadt, Germany) and VS2 contained 15% EG, 15% DMSO, 0.5 mol/l trehalose (Merck, Darmstadt, Germany) in HM. Furthermore, warming solution (WS) was divided into four groups. There were: WS1a contained 0.3 mol/l sucrose, WS1b contained 0.15 mol/l sucrose, WS2a contained 0.3 mol/l trehalose, and WS2b contained 0.15 mol/l trehalose. Apoptotic level was performed by staining the oocytes with TUNEL and propidium iodide (PI) based on Brison and Schultz method then examined under confocal microscope. The rate of apoptosis in oocytes after vitrification and warming was higher compared to the fresh control oocytes. Furthermore, the rate of apoptosis in the vitrified oocytes by sucrose media (28%) was higher compared to the vitrified oocytes by trehalose media (16%). The results of this study indicated that vitrification increased apoptosis in the vitrified oocytes related to the oocyte injury after vitrification. Moreover, the vitrification increased apoptosis more in the vitrified oocytes by sucrose media than the vitrified oocytes by trehalose media. The exposure to the 16.5% EG, 16.5% DMSO and 0.5 mol/l trehalose as cryoprotectant media decreased their viability and increased the number of DNA-fragmented nuclei in the oocytes, lesser than sucrose. Trehalose was proved to be the more suitable extracellular cryoprotectant media in oocyte vitrification based on the apoptotic level, compared to that of sucrose. A modification of cryoprotectant media as an update of oocyte vitrification consisted 0.5 mol/l trehalose concentration as extracellular cryoprotectant and combined with 16.5% EG and 16.5% DMSO as intracellular cryoprotectant has produced.

Original languageEnglish
Title of host publication2nd Biomedical Engineering�s Recent Progress in Biomaterials, Drugs Development, and Medical Devices
Subtitle of host publicationProceedings of the International Symposium of Biomedical Engineering, ISBE 2017
EditorsRadon Dhelika, Yudan Whulanza, Ghiska Ramahdita, Praswasti P.D.K. Wulan
PublisherAmerican Institute of Physics Inc.
ISBN (Electronic)9780735416253
DOIs
Publication statusPublished - 13 Feb 2018
Event2nd Biomedical Engineering's Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2017 - Bali, Indonesia
Duration: 25 Jul 201726 Jul 2017

Publication series

NameAIP Conference Proceedings
Volume1933
ISSN (Print)0094-243X
ISSN (Electronic)1551-7616

Conference

Conference2nd Biomedical Engineering's Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2017
CountryIndonesia
CityBali
Period25/07/1726/07/17

Keywords

  • apoptotic level
  • oocyte vitrification
  • sucrose
  • trehalose

Fingerprint Dive into the research topics of 'Comparison of sucrose and trehalose media modification as an update of oocyte vitrification: A study of apoptotic level'. Together they form a unique fingerprint.

Cite this