Generation of functional eggs and sperm from cryopreserved whole testes.
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ABSTRACT:
SUBMITTER: Lee S
PROVIDER: S-EPMC3562789 | biostudies-literature | 2013 Jan
REPOSITORIES: biostudies-literature
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Generation of functional eggs and sperm from cryopreserved whole testes.
Lee Seungki S Iwasaki Yoshiko Y Shikina Shinya S Yoshizaki Goro G
Proceedings of the National Academy of Sciences of the United States of America 20130114 5
The conservation of endangered fish is of critical importance. Cryobanking could provide an effective backup measure for use in conjunction with the conservation of natural populations; however, methodology for cryopreservation of fish eggs and embryos has not yet been developed. The present study established a methodology capable of deriving functional eggs and sperm from frozen type A spermatogonia (ASGs). Whole testes taken from rainbow trout were slowly frozen in a cryomedium, and the viabil ...[more]
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RNA was extracted from each frozen-thawed sperm sample and randomized into pools (4 pools for short-time samples and 3 pools for long-time samples), each of 5-6 patients. Wide-transcriptome comparison of short- and long-time pools were analyzed. Participants/materials, setting, methods The study population had a median age of 34.5 years (range: 22-46), without significant difference between the two groups (p-value = 0.147). RNA was extracted by miRNeasy Micro Kit (Qiagen), analyzed by High Sensitivity RNA on 2200 Tape Station system (Agilent), amplified by Ovation Pico WTA SystemV2 (Nugen), labeled and hybridized on GE 4x44K v2 microarrays (Agilent). The unpaired Significance Analysis of Microarray (SAM) was performed using the limma, hgug4112a.db and samr packages in R/BioConductor. 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