Title:
METHOD FOR ULTRA-LOW TEMPERATURE CRYOPRESERVATION OF SEA CUCUMBER SPERM
Document Type and Number:
WIPO Patent Application WO/2021/012763
Kind Code:
A1
Abstract:
Disclosed is a method for ultra-low temperature cryopreservation of sea cucumber sperm, which is performed according to the following steps, in sequence: taking a sea cucumber during the breeding period and wiping dry the body of the sea cucumber to remove seawater; after dissecting the abdomen, removing the complete sex gland, and soaking up the body fluid of the surface of the complete sex gland and transferring it to a sterile petri dish to cut into pieces, and filtering the seminal fluid using a 300-mesh sieve; taking the filtered seminal fluid and a cryopreservation solution according to the volume ratio of 1:1–2 and injecting them into a cryotube and mixing well, the cryopreservation solution being made by adding to each liter of distilled water 8–15 g glucose, 7–9 g sodium chloride, 0.5–0.7 g potassium chloride, 3–5 g trehalose, 0.05–0.1 g anhydrous calcium chloride, and 80–120 mL dimethyl sulfoxide of purity ≥99.7%; suspending the cryotube at a distance of 10–15 cm from the surface of liquid nitrogen for 10 minutes, then suspending the cryotube on the surface of liquid nitrogen for 5 minutes, and finally immersing the cryotube in liquid nitrogen and keeping it frozen.
Inventors:
CHANG YAQING (CN)
ZHAN YAOYAO (CN)
ZHAO TANJUN (CN)
SONG JIAN (CN)
ZHANG WEIJIE (CN)
ZHAN YAOYAO (CN)
ZHAO TANJUN (CN)
SONG JIAN (CN)
ZHANG WEIJIE (CN)
Application Number:
PCT/CN2020/090969
Publication Date:
January 28, 2021
Filing Date:
May 19, 2020
Export Citation:
Assignee:
UNIV DALIAN OCEAN (CN)
International Classes:
A01N1/02
Foreign References:
| CN110326610A | 2019-10-15 | |||
| CN104054697A | 2014-09-24 | |||
| CN107183014A | 2017-09-22 | |||
| KR20150101498A | 2015-09-04 |
Other References:
MING YU SHAO , ZHI FENG ZHANG , LI YU , JING JIE HU , KYOUNG HO KANG: "Cryopreservation of sea cucumber Apostichopus japonicus (Selenka) sperm", AQUACULUTRE RESERCH, vol. 37, no. 14, 30 July 2006 (2006-07-30), pages 1450 - 1457, XP055775534, ISSN: 1355-557X, DOI: 10.1111/j.1365-2109.2006.01581.x
Attorney, Agent or Firm:
DALIAN FEIFAN PATENT AGENCY (CN)
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