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Comparison of two culture methods during in vitro spermatogenesis of vitrified-warmed testis tissue: Organ culture vs. hanging drop culture.

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Title: Comparison of two culture methods during in vitro spermatogenesis of vitrified-warmed testis tissue: Organ culture vs. hanging drop culture.
Authors: Patra, Tanushree1 (AUTHOR), Pathak, Devendra2 (AUTHOR), Gupta, Mukesh Kumar1 (AUTHOR) guptam@nitrkl.ac.in
Source: Cryobiology. Jun2021, Vol. 100, p142-150. 9p.
Subjects: Organ culture, Tissue culture, Spermatogenesis, Testis, Cross-cultural studies, Histology
Abstract: Solid surface vitrification (SSV) is a cost effective and simple method for testis tissue preservation. Vitrified-warmed testis tissue was successfully cultured using various organ culture methods. In this study, we compared two culture methods viz. hanging drop (HD) and organ culture (OC) methods for in vitro spermatogenesis of goat testis tissue vitrified-warmed by SSV. It was observed that OC method was superior (p < 0.05) to HD method in terms of post-warming metabolic activity of testicular tissue, as measured by MTT assay on Day 7 and Day 14 of culture, respectively. The size of the tissue also played an important role in post-warming metabolic activity and viability (4 mm3: 72.7 ± 1.2% vs. 9 mm3: 62.7 ± 1.3% vs. 16 mm3: 40.5 ± 1.7%) of vitrified tissues with smaller tissue resulting in better result. The vitrification-induced ROS activity significantly decreased during their in vitro culture. Histology and scanning electron microscopy (SEM) showed the rupture of basal membrane, surface morphology and, cell loss due to vitrification. However, histology and immunohistochemistry showed the progression of in vitro spermatogenesis and formation of elongated spermatozoa in both fresh and vitrified-warmed testis tissue cultured by OC method. Taken together, our results suggest that OC method is superior to HD method for culturing goat testis tissue vitrified-warmed by SSV. • Viability of vitrified-warmed goat testicular tissue varied with the tissue size: smaller the tissue size, better the post-warming viability. • In vitro culture by organ culture method was superior to hanging drop method for culturing vitrified-warmed testis tissue. • Histology and immunohistochemistry confirmed the spermatogenic progression and sperm production in vitrified-warmed testis tissue by organ culture. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
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Abstract:Solid surface vitrification (SSV) is a cost effective and simple method for testis tissue preservation. Vitrified-warmed testis tissue was successfully cultured using various organ culture methods. In this study, we compared two culture methods viz. hanging drop (HD) and organ culture (OC) methods for in vitro spermatogenesis of goat testis tissue vitrified-warmed by SSV. It was observed that OC method was superior (p < 0.05) to HD method in terms of post-warming metabolic activity of testicular tissue, as measured by MTT assay on Day 7 and Day 14 of culture, respectively. The size of the tissue also played an important role in post-warming metabolic activity and viability (4 mm3: 72.7 ± 1.2% vs. 9 mm3: 62.7 ± 1.3% vs. 16 mm3: 40.5 ± 1.7%) of vitrified tissues with smaller tissue resulting in better result. The vitrification-induced ROS activity significantly decreased during their in vitro culture. Histology and scanning electron microscopy (SEM) showed the rupture of basal membrane, surface morphology and, cell loss due to vitrification. However, histology and immunohistochemistry showed the progression of in vitro spermatogenesis and formation of elongated spermatozoa in both fresh and vitrified-warmed testis tissue cultured by OC method. Taken together, our results suggest that OC method is superior to HD method for culturing goat testis tissue vitrified-warmed by SSV. • Viability of vitrified-warmed goat testicular tissue varied with the tissue size: smaller the tissue size, better the post-warming viability. • In vitro culture by organ culture method was superior to hanging drop method for culturing vitrified-warmed testis tissue. • Histology and immunohistochemistry confirmed the spermatogenic progression and sperm production in vitrified-warmed testis tissue by organ culture. [ABSTRACT FROM AUTHOR]
ISSN:00112240
DOI:10.1016/j.cryobiol.2021.02.006