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Retinol Improves In Vitro Differentiation of Pre-Pubertal Mouse Spermatogonial Stem Cells into Sperm during the First Wave of Spermatogenesis

View Article: PubMed Central - PubMed

ABSTRACT

Testicular tissue freezing has been proposed for fertility preservation in pre-pubertal boys. Thawed frozen testicular tissue must undergo a maturation process to restore sperm production. The purpose of the current study was to evaluate the ability of retinol to improve the in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm. Testes from pre-pubertal mice, aged 2.5 and 6.5 days post-partum, were cultured on agarose gel at a gas-liquid interphase for 34, 38 and 60 days (D) and for 16, 30 and 36 D respectively. Assessment of basal medium (BM) supplemented with retinol (RE) alone, FSH/LH alone or a combination of both, was performed. Stereological analyses and tissue lesion scoring were performed at the culture time points indicated above. Sperm production was quantified at D30 and D34 after mechanical dissection of the testicular tissues. FSH/LH significantly increased the percentage of round spermatids at D30 and D38, when compared to BM alone. However, RE significantly increased the percentages of round but also elongated spermatids at D30 and D34. Moreover, RE significantly increased the number of spermatozoa per milligram of tissue at D30 and D34 when compared to BM. Therefore, RE improved the in vitro production of spermatids and spermatozoa from pre-pubertal SSCs during the first wave of spermatogenesis. The use of RE could be a useful tool for in vitro spermatogenesis from pre-pubertal human testicular tissue.

No MeSH data available.


Related in: MedlinePlus

Seminiferous tubule growth (A1, A2), intra-tubular cell density (A3, A4), germ/Sertoli cells ratio (A5 and A6) and intra-tubular cell proliferation (B1, B2) during organotypic culture of prepubertal mice testes under different culture conditions.The results are presented as the mean ± s.e.m., with n = 4. Asterisk indicates a statistically significant difference (p<0.05). Footnotes: BM: Basal Medium, D: Day, dpp: day post-partum, FSH: Follicle Stimulating Hormone, LH: Luteinizing Hormone, n: Number of mice testes used in each condition, PCNA: Proliferating Cell Nuclear Antigen, RE: Retinol, s.e.m.: Standard Error of the Mean
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pone.0116660.g002: Seminiferous tubule growth (A1, A2), intra-tubular cell density (A3, A4), germ/Sertoli cells ratio (A5 and A6) and intra-tubular cell proliferation (B1, B2) during organotypic culture of prepubertal mice testes under different culture conditions.The results are presented as the mean ± s.e.m., with n = 4. Asterisk indicates a statistically significant difference (p<0.05). Footnotes: BM: Basal Medium, D: Day, dpp: day post-partum, FSH: Follicle Stimulating Hormone, LH: Luteinizing Hormone, n: Number of mice testes used in each condition, PCNA: Proliferating Cell Nuclear Antigen, RE: Retinol, s.e.m.: Standard Error of the Mean

Mentions: The seminiferous tubule surface (Fig. 2A1) increased significantly during the culture for FSH/LH (p = 0.04) and RE (p = 0.0046) as observed in vivo (p = 0.0046), even if this surface was significantly higher in the corresponding in vivo controls than under in vitro conditions (p = 0.01). Moreover, the greatest seminiferous tubule areas were obtained with FSH/LH when compared with BM and RE at D38 (p = 0.02 and p = 0.01 respectively) and D60 (p = 0.01). However, seminiferous tubule surface was similar at D38 and D60 between BM and RE.


Retinol Improves In Vitro Differentiation of Pre-Pubertal Mouse Spermatogonial Stem Cells into Sperm during the First Wave of Spermatogenesis
Seminiferous tubule growth (A1, A2), intra-tubular cell density (A3, A4), germ/Sertoli cells ratio (A5 and A6) and intra-tubular cell proliferation (B1, B2) during organotypic culture of prepubertal mice testes under different culture conditions.The results are presented as the mean ± s.e.m., with n = 4. Asterisk indicates a statistically significant difference (p<0.05). Footnotes: BM: Basal Medium, D: Day, dpp: day post-partum, FSH: Follicle Stimulating Hormone, LH: Luteinizing Hormone, n: Number of mice testes used in each condition, PCNA: Proliferating Cell Nuclear Antigen, RE: Retinol, s.e.m.: Standard Error of the Mean
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340963&req=5

pone.0116660.g002: Seminiferous tubule growth (A1, A2), intra-tubular cell density (A3, A4), germ/Sertoli cells ratio (A5 and A6) and intra-tubular cell proliferation (B1, B2) during organotypic culture of prepubertal mice testes under different culture conditions.The results are presented as the mean ± s.e.m., with n = 4. Asterisk indicates a statistically significant difference (p<0.05). Footnotes: BM: Basal Medium, D: Day, dpp: day post-partum, FSH: Follicle Stimulating Hormone, LH: Luteinizing Hormone, n: Number of mice testes used in each condition, PCNA: Proliferating Cell Nuclear Antigen, RE: Retinol, s.e.m.: Standard Error of the Mean
Mentions: The seminiferous tubule surface (Fig. 2A1) increased significantly during the culture for FSH/LH (p = 0.04) and RE (p = 0.0046) as observed in vivo (p = 0.0046), even if this surface was significantly higher in the corresponding in vivo controls than under in vitro conditions (p = 0.01). Moreover, the greatest seminiferous tubule areas were obtained with FSH/LH when compared with BM and RE at D38 (p = 0.02 and p = 0.01 respectively) and D60 (p = 0.01). However, seminiferous tubule surface was similar at D38 and D60 between BM and RE.

View Article: PubMed Central - PubMed

ABSTRACT

Testicular tissue freezing has been proposed for fertility preservation in pre-pubertal boys. Thawed frozen testicular tissue must undergo a maturation process to restore sperm production. The purpose of the current study was to evaluate the ability of retinol to improve the in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm. Testes from pre-pubertal mice, aged 2.5 and 6.5 days post-partum, were cultured on agarose gel at a gas-liquid interphase for 34, 38 and 60 days (D) and for 16, 30 and 36 D respectively. Assessment of basal medium (BM) supplemented with retinol (RE) alone, FSH/LH alone or a combination of both, was performed. Stereological analyses and tissue lesion scoring were performed at the culture time points indicated above. Sperm production was quantified at D30 and D34 after mechanical dissection of the testicular tissues. FSH/LH significantly increased the percentage of round spermatids at D30 and D38, when compared to BM alone. However, RE significantly increased the percentages of round but also elongated spermatids at D30 and D34. Moreover, RE significantly increased the number of spermatozoa per milligram of tissue at D30 and D34 when compared to BM. Therefore, RE improved the in vitro production of spermatids and spermatozoa from pre-pubertal SSCs during the first wave of spermatogenesis. The use of RE could be a useful tool for in vitro spermatogenesis from pre-pubertal human testicular tissue.

No MeSH data available.


Related in: MedlinePlus