Cell survival after cryopreservation of dissociated testicular cells from feline species |
| |
| Institution: | 1. Guangdong Province Key Laboratory of Reproductive Medicine, the First Affiliated Hospital and State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China;2. The Key Laboratory for Mammalian Reproductive Biology and Biotechnology, Ministry of Education, Inner Mongolia University, Hohhot, China;1. Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran;2. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;3. Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran;4. Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Science, Shahrekord, Iran;5. Urology Research Center, Sina Hospital, TehranUniversity of Medical Sciences, Tehran, Iran;6. Gametogenesis Research Center, Kashan University of Medical Sciences, Kashan, Iran;7. School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran;8. Infertility and Reproductive Medicine, Department of Obstetrics and Gynecology, Shiraz University of Medicine Sciences, Shiraz, Iran;9. Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran;1. Dipartimento di Scienze Veterinarie per la Salute, la Produzione Animale e la Sicurezza Alimentare, Università degli Studi di Milano, Milano Italy;2. Ospedale Grandi Animali, Università degli Studi di Milano, Lodi, Italy;3. Dipartimento di Scienze Veterinarie e Sanità Pubblica, Università degli Studi di Milano, Milano, Italy |
| |
| Abstract: | Testicular cell suspension (TCS) can be cryopreserved for male germ-line preservation and fertility restoration. We aimed to validate a cryopreservation protocol for TCS of domestic cat to be applied in endangered felids species. Testis tissue from adult domestic cats was enzymatically dissociated and spermatogenic cells were enriched. The resulting TCS was diluted in 7.5% or 15% Me2SO based medium. Slow and fast freezing methods were tested. We examined the effects of freezing approaches using two combinations of fluorescent dyes: Calcein-AM with Propidium iodide (C/PI) and SYBR14 with Propidium iodide (S/PI). Ploidy analysis of domestic cat fresh TCS revealed that the majority of testicular cells were haploid cells. Based on microscopic observation, two size populations (12.3 ± 2.3 μm and 20.5 ± 4 μm in diameter) were identified and presumed to be mainly spermatids and spermatocytes, respectively. Both evaluation methods proved higher viability of aggregated cells before and after cryopreservation compared with single cells, and superiority of low concentration of Me2SO (7.5%) in association with slow freezing to preserve viability of testicular cells. However, S/PI resulted in a more precise evaluation compared with the C/PI method. The combination of 7.5% Me2SO-based medium with slow freezing yielded post thaw viability of S/PI labeled aggregated (49.8 ± 20%) and single cells (31.5 ± 8.1%). Comparable results were achieved using testes of a Cheetah and an Asiatic golden cat. In conclusion, TCS from domestic cat can be successfully cryopreserved and has the potential to support fertility restoration of endangered felids species. |
| |
| Keywords: | Felids Testicular cells Me2SO Cryopreservation |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
