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Method for transplanting consubstantial cell nucleus

A technology of somatic cell nuclear transfer and nucleus, which is applied in the field of biological homogeneous cell nuclear transfer, can solve the problems of DNA mismatch, abnormal embryo development, and low efficiency of cloning technology, and achieve the effect of improving cloning efficiency and development

Active Publication Date: 2004-12-29
SHANGHAI INST OF MEDICAL GENETICS SHANGHAI CHILDRENS HOSPITAL
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Problems solved by technology

[0005] In summary, one of the main reasons for the inefficiency of cloning technology is that the nucleus of the donor cell does not match the DNA of the recipient cytoplasm, mainly manifested in the incompatibility of the nuclear genome and the cytoplasmic genome
The reason is that the nucleus of the donor cell and the cytoplasm of the recipient cell come from two animal individuals with different genetic backgrounds, which leads to the disorder of the energy metabolism of the reconstituted embryo formed by somatic cell nuclear transfer, resulting in abnormal embryo development and cloned embryos. high mortality

Method used

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Examples

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Embodiment 1

[0026] Example 1 Allogeneic somatic cell nuclei and quality clone bovine

[0027] 1. Prepare donor cells, take skin fibroblasts or cumulus cells of young cattle with clear genetic background according to known methods, and culture them with D-MEM / F-12 with 10% serum, and use 1-5 generations And 0.5% serum starved cells for 2-4 days for routine cloning.

[0028] 2. Prepare recipient oocytes. Under B-mode ultrasound monitoring, collect eggs from the same individual as the donor cells, disinfect and anesthetize the cow, and insert the B-ultrasound probe with egg collection needle into the vagina gently. The fornix pulls the ovary to the end of the probe through the rectal wall, and advances the egg collection needle. At the same time, the foot switch controls the vacuum pump to suck the follicle fluid, and the pressure is about 95mmHg. Wash with DPBS containing 3% BSA and 2u / ml heparin Rinse the egg collection needle and catheter, and put into the collection tube. Pour the coll...

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Abstract

The present invention belongs to the field of bioengineering technology, and is especially biological consubstantial cell nucleus grafting and cloning technology. By means of biological cloning technology, the present invention takes consubstantial nucleus donor cell and denucleated egg mother cell with clear genetic background for consubstantial cell nucleus grafting to prepare cloned animal. The present invention has relatively high matching between the donor nucleus DNA and acceptor cytoplasm and the electronic compatibility inside nuclear substance, and can improve the development of cloned embryo and raise cloning efficiency. The said technology may be used in cloning animal to obtain high quality livestock.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a biosimilar cell nucleus transplantation technology. Background technique [0002] In recent years, bioengineering cloning technology has been developed vigorously. It has been reported that animal cloning technology is to transplant the donor somatic cell nucleus into the cytoplasm of the allogeneic recipient enucleated oocyte through microsurgery and cell fusion to reconstitute the embryo ( reconstituted embryos). The reconstructed embryos are cultured to develop into blastocysts, and then the blastocysts are transplanted into the uterus of the host mother who is in estrus synchronously, and the animals are delivered after they mature. The animals are called cloned animals, and the technology is called somatic cell cloning. Also known as somatic cell nuclear transfer. [0003] Mammalian somatic cell cloning technology has made great breakthroughs after sev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/16
Inventor 曾溢滔黄淑帧杨哓煜李华
Owner SHANGHAI INST OF MEDICAL GENETICS SHANGHAI CHILDRENS HOSPITAL
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