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Cryopreservation of sperm

A sperm, low-speed technology, applied in the preservation of human or animal bodies, microorganisms, biochemical equipment and methods, etc., can solve problems such as cell damage

Inactive Publication Date: 2003-10-22
GTC BIOTHERAPEUTICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the freezing and thawing process can cause dehydration of the cells leading to cell damage

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Thirty-two billy goats consisting of three breeds (Alpine, Saanen and Toggenburg) aged 13 days to 7 years were used. The animals were treated according to protocols approved by the Institutional Animal Care and Use Committee (IACUC) and in accordance with the terms set forth in the Animal Welfare Act (AWA). Animals were euthanized by intravenous barbiturate overdose. The testes were removed from the scrotum within about 5 to about 10 minutes, and these testes were then placed in an incubator at 38°C. Testicles are processed individuals. The capsule was peeled off using a sterile scalpel to expose the caudal epididymis. A small lateral incision is made along the caudal epididymis to open the coiled tubules. Gently compress the tail, thereby producing a few small drops of semen. The few drops of semen were transferred with a pipette to a supplement solution (Continental Plastic Corp., Delavan WI) equilibrated with 20% v / v egg yolk per 100 ml (standard-pathogen-free, S...

Embodiment 2

[0091] Sperm from transgenic male rabbits were collected using an artificial vagina. Samples were diluted with glycerol-free Part A make-up solution and placed in thermos flasks at approximately 37°C in the laboratory. Samples were processed as in Example 1 with the following exceptions. Twenty million sperm are loaded into each thin tube. After cooling the sample to 4 °C, add the Part B replenishment solution containing glycerol. The samples were then frozen to -80° C. specifically as described in Example 1 and stored in liquid nitrogen for 1 month. Four Dutch rabbits were synchronized by administering the hormones follicle-stimulating hormone and human chorionic gonadotropin. Sperm samples were thawed by placing them in a water bath at 37°C for 90 seconds. The samples were then used for artificial insemination of synchronous estrus female rabbits. The two female rabbits inseminated with the samples gave birth to a total of eleven offspring.

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PUM

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Abstract

The invention features methods of cryopreserving sperm. The methods include providing a sample of sperm; cooling the sperm to a first temperature; maintaining the sperm sample at the first temperature; cooling the sperm to a second temperature; maintaining the sperm at the second temperature; and storing the sperm at a third temperature. These methods can be used to cryopreserve sperm from mammals, e.g., transgenic mammals and can be to preserve sperm for subsequent artificial insemination or in vitro fertilization.

Description

Background technique [0001] The skill of modifying animal genomes through transgenic technology has opened up new avenues for medical applications. At present, the expression of biopharmaceutical proteins through the mammary glands of farm animals makes it possible to produce large quantities of valuable therapeutic proteins at low cost (Houdebine (1995) Reprod.Nutr.Dev.35: 609-617; Maga et al. (1995) Biotechnology ( Bio / Technology), 13:1452-1457; Echelard (1996) Curr.Op. (Biotechnol.7:536-540; Young et al. (1997) Biopharmaceuticals (BioPharm) 10:34-38). Although the 15 best The total sales of biopharmaceuticals in 1996 have reached 7.5 billion US dollars, but this value is expected to continue to increase in the future (Med AdNews 16:30). Whether it is due to the demand for high unit doses, The number of administrations or the existence of a large patient population results in a large demand for proteins for which transgenic technology is applicable and beneficial, for compl...

Claims

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Application Information

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IPC IPC(8): A01K67/02A01N1/02C12N5/07C12N5/076
CPCA01N1/0284A01N1/02A01N1/0221
Inventor W·A·伽温S·M·布拉施C·A·卡姆索D·T·迈利坎
Owner GTC BIOTHERAPEUTICS INC