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Refrigeration method for preserving embryo and oocyte by vitrification of glasscapillary

A vitrification freezing, capillary glass tube technology, applied in the field of embryo and oocyte preservation, can solve the problems of osmotic pressure damage, chemical toxicity damage, difficult operation, etc., and achieve low cost, simple operation and high survival rate. Effect

Inactive Publication Date: 2007-07-25
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current problem with vitrification is that high-concentration vitrification solution will inevitably cause chemical toxicity and osmotic damage to embryos and oocytes.
However, the problem with this method is that it is difficult to operate

Method used

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  • Refrigeration method for preserving embryo and oocyte by vitrification of glasscapillary

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Configuration of frozen liquid:

[0031] Base solution: add 2.5ml fetal calf serum (V / V) and 0.1ml gentamicin solution (V / V) to 10ml tissue and cell culture medium (TCM-199 solution);

[0032] Vitrification solution I: add 1.2ml ethylene glycol (V / V) and 1.3ml dimethyl sulfoxide (V / V) to 10ml base solution;

[0033] Vitrification solution II: add 3.2ml ethylene glycol (V / V) and 3.4ml dimethyl sulfoxide (V / V) to 10ml base solution;

[0034] (two), freezing:

[0035]Using a four-hole plate, place 0.8 ml of base solution, vitrification solution I and vitrification solution II in the first three wells of the four-hole plate; place mature bovine oocytes to be frozen in the base solution first; Use a pipette to put 3 bovine mature oocytes into vitrification solution I and equilibrate for 1.5 minutes; then transfer the bovine mature oocytes to vitrification solution II, and then use a pipette to aspirate Transfer the oocytes together with 5 microliters of vitrification ...

Embodiment 2

[0043] (1) Configuration of frozen liquid:

[0044] Base solution: add 2ml fetal bovine serum (V / V) and 0.15ml gentamicin solution (V / V) to 10ml tissue and cell culture medium (TCM-199 solution);

[0045] Vitrification solution I: add 1.5ml ethylene glycol (V / V) and 1.2ml dimethyl sulfoxide (V / V) to 10ml base solution;

[0046] Vitrification solution II: add 3.5ml ethylene glycol (V / V) and 3ml dimethyl sulfoxide (V / V) to 10ml base solution;

[0047] (two), freezing:

[0048] Use a four-well plate, place 0.8 ml of base solution, vitrification solution I, and vitrification solution II in the first three wells of the four-well plate; put the bovine embryos to be frozen in the base solution first; use a pipette Equilibrate 2 bovine embryos in vitrification solution I for 1.5 minutes; then transfer the bovine embryos to vitrification solution II; then pipette the bovine embryos together with 6 microliters of vitrification solution II is transferred to the blank hole; the capilla...

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Abstract

This invention discloses a method for preserving embryo and oocyte through vitrification cryopreservation by utilizing capillary glass tube. The method utilizes high borosilicate capillary glass tube, and comprises: (1) preparing freezing liquid; (2) freezing; (3) preparing thawing liquid; (4) thawing. The method utilizes high borosilicate capillary glass tube to preserve embryo and oocyte, and has such advantages as no need for expensive devices, low cost, easy operation, good vitrification cryopreservation effect, and high survival rate after thawing.

Description

technical field [0001] The invention relates to a method for preserving embryos and oocytes, in particular to a method for vitrifying and freezing preservation of embryos and oocytes with capillary glass tubes. Background technique [0002] As a newly developed biotechnology, vitrification technology has many potential applications. Applying this technology to cryopreservation of embryos and oocytes can save early embryos of fine animals and a large number of oocytes in discarded ovaries, and establish "embryo banks" and "egg banks" for embryo transfer, in vitro The development of various embryonic biotechnologies such as fertilization, nuclear transfer, and transgenics provides sufficient materials and removes time and space constraints; it can also provide the possibility for the preservation of genetic resources of rare and endangered animals. Therefore, it has broad application prospects in the field of developmental reproduction research. [0003] Vitrification refers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06A01N1/02
Inventor 谭秀文黄金明游伟刘晓牧吴乃科
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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