Effects of different cryopreservation type on testicular tissue of mice: histological changes study

Abstract

Cryopreservation is a process where cells or whole tissues are preserved by cooling to low sub-zero temperatures, such as (typically) -80°C or -196°C and this process use extensively in programs of in vitro fertilization (IVF). This study is an attempt to evaluate DNA fragmentation in the cell after cryopreservation/thawing cycle when using different types of cryoprotectant (CPA). Now with recent improvement of assisted reproductive technologies and possibility of using testicular spermatozoa or epididymal spermatozoa at in vitro fertilization by intracytoplasmic sperm injection (ICSI), the cryopreservation of testicular tissue become widely used in male who were treated by chemotherapy or radiotherapy because these therapeutic applications can lead to sterility. This study is an attempt to investigate the changes that occur in mice testicular tissue after cryopreservation/thawing cycle as a model for human being after cryopreservation/thawing cycle when using different types of cryoprotectant (CPA). Sixty mature fertile male mice were used in the current study, the mean age of these mice were ten weeks. Fifteen of them were considered as control group and the rest (forty five) as cryopreserved group, this group was divided into three subgroup according to the type of cryoprotectant (glycerol, 1, 2 propanediol and dimethylsulfoxide), each subgroup composed from fifteen mice. The testes were cryopreserved for six weeks then histological evaluation was done by paraffin section. The microscopically observation of slides obtained from cryopreserved testis differs according to the type of cryoprotectant, in the dimethylsulfoxide group the tissue appeared well-preserved cell and mild changes in the interstitial tissue, but the glycerol group shown moderate changes in testicular tissue after cryopreservation such as; rupture of the stroma, detachment the cells form basement membrane in the seminiferous tubules, gaps in germinal epithelium and folding in the lamina propria, while in the 1,2 propanediol group the tissue shows sever changes such as distraction the interstitial tissue , necrotic cells in the seminiferous tubule and difficult to recognize the cells in the seminiferous tubule. From the results of the present study, it was concluded that there was alteration in testis histology in the testicular tissue after cryopreservation /thawing cycle differs according to the type of cryoprotectant and the Dimethylsulfoxide cryoprotectant provides good protection to the testicular tissue and DNA in cryopreservation /thawing cycle than glycerol and 1, 2 propanediol (1, 2 propanediol was not preferable in testicular tissue cryopreservation).

Keywords: Cryopreservation; Testicular Tissue; Cryoprotectant

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