Effective cryopreservation protocol for preservation of male primate (Macaca fascicularis) prepubertal fertility
- Authors
- Jung, Sang-Eun; Ahn, Jin Seop; Kim, Yong-Hee; Kim, Bang-Jin; Won, Jong-Hyun; Ryu, Buom-Yong
- Issue Date
- Dec-2020
- Publisher
- Elsevier Ltd
- Keywords
- Cryoprotectants; Infertility; Primate spermatogonial stem cells; Testicular tissue freezing; Trehalose; Xenotransplantation
- Citation
- Reproductive BioMedicine Online, v.41, no.6, pp 1070 - 1083
- Pages
- 14
- Journal Title
- Reproductive BioMedicine Online
- Volume
- 41
- Number
- 6
- Start Page
- 1070
- End Page
- 1083
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/52056
- DOI
- 10.1016/j.rbmo.2020.07.016
- ISSN
- 1472-6483
1472-6491
- Abstract
- Research question: Can specimen types (cells versus tissues) and additive cryoprotectant agents contribute to efficient cryopreservation of primate spermatogonial stem cells (SSC)? Design: Testicular tissues or cells from four prepubertal monkeys were used in this study. The freezing efficacy of testicular tissue was compared with cell suspensions using conventional freezing media (1.4 mol/l dimethyl sulfoxide [DMSO]) and the efficacy of cryoprotectant additives (1.4 mol/l DMSO combined with trehalose 200 mmol/l, hypotaurine 14 mmol/l, necrostatin-1 50 µmol/l or melatonin 100 µmol/l) was evaluated in testicular tissue freezing. Results: The survival rate (46.0 ± 4.8% versus 33.7 ± 6.0%; P = 0.0286) and number of recovered cells (5.0 ± 1.5 × 106 cells/g versus 0.7 ± 0.8 × 106 cells/g; P = 0.0286) were significantly higher in frozen tissues than in frozen cell suspensions. After tissue freezing, a higher number of recovered PGP9.5+ cells were observed with 200 mmol/l trehalose treatment than in DMSO controls (2.4 ± 0.6 × 106 cells/g versus 1.1 ± 0.3 × 106 cells/g; P = 0.0164). Normal establishment of donor-derived colony was observed in SSC after tissue freezing with 200 mmol/l trehalose. Conclusions: Testicular tissue freezing is more effective than single cell suspension freezing for higher recovery of undifferentiated spermatogonia. Moreover, it was verified that slow freezing using 200 mmol/l trehalose, 1.4 mol/l DMSO and 10% KnockOut™ Serum Replacement in Dulbecco's phosphate-buffered saline is an effective cryopreservation protocol for primate testicular tissue. © 2020 Reproductive Healthcare Ltd.
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