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Sheep placenta in vitro culture fluid, and its compounding method and sheep placenta in vitro culture method

An embryo culture medium and in vitro culture technology, which is applied to the sheep embryo in vitro culture medium and its preparation, and the field of livestock embryo culture, can solve the problem of low success rate of transplantation, achieve low economic cost, improve the in vitro development rate of half embryos after cutting, The effect of facilitating the promotion of the application

Inactive Publication Date: 2007-06-13
新昌县大东种畜发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem to be solved and the technical task proposed by the present invention are to overcome the defect that the transplantation success rate of the existing embryo culture solution is not high, and provide a kind of sheep embryo in vitro culture solution that can also be used as a transplantation solution; Preparation method of embryo in vitro culture solution; thirdly, a method for in vitro culture of sheep embryo is provided

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) Extract velvet extract

[0019] Descale and remove the sika velvet antler, fry the velvet antler with boiling water in an electric kettle, and bake the velvet antler in a far-infrared oven at 50-60°C for 2-6 hours. Use a semi-automatic slicer to cut into 0.35-0.50 mm thick slices, add to the pulverizer to crush the velvet antler into 400-mesh particles, boil in water for 2-4 hours, filter, discard the precipitate, and dry the supernatant with a freeze dryer for 10 ~25 hours, made into freeze-dried powder, weighed and packed separately.

[0020] (2) Making culture medium containing velvet antler

[0021] Add 1 mg of velvet antler freeze-dried powder and 8 ml of calf serum (FCS) into the graduated cylinder, and then slowly add phosphate buffered saline (PBS) to the cylinder to 100 ml. The salt buffer solution is prepared as an embryo culture solution containing 10 μg / mL of deer antler extract and 8% (volume percentage) of calf serum. (Remarks: Phosphate buffer solu...

Embodiment 2

[0032] (1) Extract velvet extract

[0033] Descale and remove the sika velvet antler, fry and boil the velvet antler in an electric kettle, and bake the velvet antler in a far-infrared oven at 50-60°C for 2-6 hours. Use a semi-automatic slicer to cut into 0.35-0.50 mm thick slices, add to the pulverizer to crush the velvet antler into 400-mesh particles, boil in water for 2-4 hours, filter, discard the precipitate, and dry the supernatant with a freeze dryer for 10 ~25 hours, made into freeze-dried powder, weighed and packed separately.

[0034] (2) Making culture medium containing velvet antler

[0035] Add 3 mg of velvet antler freeze-dried powder and 8 ml of calf serum (FCS) into the graduated cylinder, and then slowly add phosphate buffered saline (PBS) to the cylinder to 100 ml, during which the velvet antler freeze-dried powder and calf serum are fully dissolved in phosphoric acid The salt buffer solution is prepared as an embryo culture solution containing 30 μg / mL of...

Embodiment 3

[0047] (1) Extract velvet extract

[0048] Same as embodiment one.

[0049] (2) Making culture medium containing velvet antler

[0050] Add 1 mg of velvet antler freeze-dried powder and 20 ml of calf serum (FCS) into the graduated cylinder, and then slowly add phosphate buffered saline (PBS) to the cylinder to 100 ml. The salt buffer solution is prepared as an embryo culture solution containing 10 μg / mL of deer antler extract and 20% (volume percentage) of calf serum.

[0051] (3) In vitro culture of goat whole embryos and half embryos

[0052] Whole embryo culture: Two hours before the culture, in a 6cm petri dish, use a 100μl pipette to make the embryo culture solution containing 10μg / mL of velvet antler extract and 20% (volume percentage) of calf serum to a volume of 50~ 100 μL culture droplet was preheated in a 38-39°C incubator. For Boer goats pregnant for 5-6 days, 200 embryos at the mulberry stage were obtained by operation. After washing three times with PBS contai...

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PUM

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Abstract

This invention belongs to animal embryo culture technology, and disclosed culture medium of sheep embryos in vitro containing velvet extracts, and its preparation methods and in vitro embryo cultivation methods, It is primarily about extraction from the velvet antlers, addition of conventional embryo culture fluid, and for cultivation of whole sheep embryo and cut embryo in vitro, and using the medium for transplantation of whole sheep embryo. The problems about the lower development rate of whole sheep embryo in vitro, embryo transfer pregnancy rates, development rate of the cut embryos in vitro were solved with the application of Chinese medicine velvet extract in field of sheep embryos in vitro. Its economy is lower cost and it is facilitated to application in production.

Description

technical field [0001] The invention discloses a sheep embryo in vitro culture solution containing deer antler extract and a preparation method thereof, belonging to the technical field of livestock embryo culture methods. Background technique [0002] Embryo transfer is a new breeding technology applied to mammals, and it is an important part of embryo engineering. It has become a relatively mature biotechnology. Through embryo transfer, excellent female animals can be used to maximize their performance in variety improvement and breeding. good effect. The in vitro culture of embryos is usually the pre-procedure of successful embryo transfer, such as through the in vitro culture of embryos can promote the development of embryos and seek a suitable transplant culture medium. Embryo cutting is a commonly used auxiliary technology for embryo transfer. By cutting early embryos, on the one hand, the number of embryos can be increased, and on the other hand, it can provide mater...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/073
Inventor 徐雪明俞颂东王争光王锡炉张锐
Owner 新昌县大东种畜发展有限公司
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