Biopolymer gels as a basis of cryoprotective medium for testicular tissue of rats

Волкова, Н. А.; Yukhta, Mariia; Goltsev, Anatoliy

doi:10.1007/s10561-018-9740-z

Cited by 13 publications

(7 citation statements)

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“…This was confirmed in mouse immature testicular tissues encapsulated in a fibrin hydrogel [28]. Fibrin gel could serve as an integral component of the cryopreservation medium, which effectively contributed to the preservation of the average density of germinative cells and their metabolic activity (total lactate dehydrogenase activity) [28]. In addition to vitrification, fibrin encapsulation supported post-thawed follicle viability compared to non-encapsulated samples.…”

Section: Discussionmentioning

confidence: 87%

“…Therefore, hydrogel-based encapsulation has been considered an effective approach to preserve cells during cryopreservation [24]. This was confirmed in mouse immature testicular tissues encapsulated in a fibrin hydrogel [28]. Fibrin gel could serve as an integral component of the cryopreservation medium, which effectively contributed to the preservation of the average density of germinative cells and their metabolic activity (total lactate dehydrogenase activity) [28].…”

Section: Discussionmentioning

confidence: 99%

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Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model

Thuwanut

Comizzoli

Pimpin

et al. 2021

Clin Exp Reprod Med

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Objective: Using domestic cats as a biomedical research model for fertility preservation, the present study aimed to characterize the influences of ovarian tissue encapsulation in biodegradable hydrogel matrix (fibrinogen/thrombin) on resilience to cryopreservation, and static versus non-static culture systems following ovarian tissue encapsulation and cryopreservation on follicle quality.Methods: In experiment I, ovarian tissues (n=21 animals; 567 ovarian fragments) were assigned to controls or hydrogel encapsulation with 5 or 10 mg/mL fibrinogen (5 or 10 FG). Following cryopreservation (slow freezing or vitrification), follicle viability, morphology, density, and key protein phosphorylation were assessed. In experiment II (based on the findings from experiment I), ovarian tissues (n=10 animals; 270 ovarian fragments) were encapsulated with 10 FG, cryopreserved, and in vitro cultured under static or non-static systems for 7 days followed by similar follicle quality assessments. Results: In experiment I, the combination of 10 FG encapsulation/slow freezing led to greater post-thawed follicle quality than in the control group, as shown by follicle viability (66.9%±2.2% vs. 61.5%±3.1%), normal follicle morphology (62.2%±2.1% vs. 55.2%±3.5%), and the relative band intensity of vascular endothelial growth factor protein phosphorylation (0.58±0.06 vs. 0.42±0.09). Experiment II demonstrated that hydrogel encapsulation promoted follicle survival and maintenance of follicle development regardless of the culture system when compared to fresh controls.Conclusion: These results provide a better understanding of the role of hydrogel encapsulation and culture systems in ovarian tissue cryopreservation and follicle quality outcomes using an animal model, paving the way for optimized approaches to human fertility preservation.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model

Thuwanut

Comizzoli

Pimpin

et al. 2021

Clin Exp Reprod Med

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“…no. Z760951, Sigma-Aldrich, St. Louis, USA), where they were exposed to the cryoprotective medium for 30 min at 4°C [8]. Then, cryotubes with samples were cooled in vapors of liquid nitrogen for 40 min down to -70°C and transferred to liquid nitrogen (-196°C) [9].…”

Section: Cryopreservationmentioning

confidence: 99%

The Use of Fullerene C60 to Preserve Testicular Tissue after Cryopreservation

Волкова

Yukhta

Goltsev

2021

Journal of Nanomaterials

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Autologous transplantation of cryopreserved fragments of an immature testis is an actively developing approach to save fertility in patients facing a gonadotoxic therapy. The use of bioavailable fullerene C60 as a powerful antioxidant opens up a new potential for the prevention and correction of ischemic-reperfusion pathological processes in tissues including those associated with freezing-thawing procedure. In this work, we aimed to study the antioxidant status, apoptotic/necrotic processes, and morphological characteristics of cryopreserved fragments of the seminiferous tubules of testis (CrFSTT) of immature rats after incubation in media with different concentrations of fullerene C60 (10, 15, and 20 μg/mL). Our results indicated that the addition of C60 in a concentration of 15 μg/mL decreased ROS production, cytochrom C release, and degree of histological damage of spermatogenic epithelium as well as increased the activity of the mitochondria, antioxidant defense system, and cell density in histological sections of CrFSTT compared to the control. Fullerene C60 at investigated concentrations did not impact significantly on apoptosis in cells of CrFSTT but, after incubation with 15 μg/mL C60, a percentage of living cells was 1.2-fold higher and a value of necrotic ones in this group was 1.6-fold lower than the control samples ( p < 0.05 ). Relative amount of cells of the spermatogonia germ layer did not differ between the studied concentrations. The general analysis of obtained data showed that the C60 addition in the concentration of 15 μg/mL was the most optimal for the rehabilitation of CrFSTT. The results can be used for the development of an effective rehabilitation medium for the cryopreserved testicular tissue.

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“…Our previous studies have shown that the use of biopolymers at the exposure stage reduced the toxic effect of DMSO and glycerol on spermatogenic epithelial cells of prepubertal rats' testis and increased the exposure time [21,22].…”

mentioning

confidence: 99%

“…The tissue fragments were transferred into 2-mL cryovials (Nunc, USA). The exposure of the rats' seminiferous tubules to the media was 30 minutes at 4 °C [21,22]. Cryopreservation was carried out using a freezer ZP-10 according to the program: cooling from 20 °С to -8 °С at a rate of 1 °С/min; stop for 10 minutes at -8 °C; cooling to -70 °C at a rate of 10 °C/min; transferring to liquid nitrogen (-196 °C).…”

mentioning

confidence: 99%

The effectiveness of biopolymers application for cryopreservation of the fragments of convoluted seminiferous tubules of prepubertal rat’s testis

Волкова¹,

Yukhta²,

Chernyschenko³

et al. 2019

JCOT

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Today, transplantation of cryopreserved fragments of immature testis is a non-alternative method of preserving the fertility of pre-adolescent patients who undergo cytotoxic therapy.The purpose of the study is to compare the effectiveness of the use of biopolymers (bovine serum albumin and fibrin gel) as the bases of cryoprotective media at low temperature preservation of the fragments of the convoluted seminiferous tubules of prepubertal rats’ testis.Materials and methods. Convoluted seminiferous tubules of prepubertal rats’ testis (75 ± 3 mg and 6-8 mm3), after a 30-minute exposure at 4 °C in cryoprotective media based on Hanks’ solution with 50 g/L of bovine serum albumin (BSA) or fibrin gel (FG) supplemented with 0.6 M DMSO or 0.7 M glycerol, were cryopreserved according to the program: 1 °C/min to -8 °C; stop for 10 minutes; 10 °C/min to -70 °C; stored in liquid nitrogen. After thawing, the histological structure was evaluated and the metabolic activity of the spermatogenic epithelium cells was determined.Results. According to the results of the histological study, there was a positive tendency of FG application, which had the maximum expressiveness in combination with 0,7 M glycerol. In this case, 68.8 ± 15.7 % of cell nuclei remained morphologically intact, and changes in the spermatogenic epithelium were slightly pronounced. The metabolic activity of the rats’ seminiferous convoluted tubules cells after freezing and thawing remained at a significantly higher level when using FG in combination with 0.6 M DMSO than with 0.7 M glycerol compared to the corresponding cryoprotectant based on the Hanks’ solution with BSA.Conclusions. The use of fibrin gel in the protocols of cryopreservation provide to preserve the histological structure and metabolic activity of the spermatogenic epithelium.

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Biopolymer gels as a basis of cryoprotective medium for testicular tissue of rats

Cited by 13 publications

References 35 publications

Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model

Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model

The Use of Fullerene C60 to Preserve Testicular Tissue after Cryopreservation

The effectiveness of biopolymers application for cryopreservation of the fragments of convoluted seminiferous tubules of prepubertal rat’s testis

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