Method for removing mammal oocyte nucleus through using wide-internal-diameter oocyte holding tube
A technology for oocytes and mammals, which is applied in the field of removing oocyte nuclei and removing mammalian oocyte nuclei with a wide inner diameter holding tube. problem, to achieve the effect of reducing the steps of enucleation, shortening the operation time of enucleation, and easy operation
Inactive Publication Date: 2013-03-27
ZHEJIANG CELLPRO BIOTECH
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However, micromanipulation such as extrusion strength is difficult to grasp, and the effect of enucleation is often unsatisfactory, or a special egg-holding tube is required, which increases the difficulty of the experiment.
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[0022] The preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, so as to define the protection scope of the present invention more clearly.
[0023] see figure 2 , prepare wide inner diameter (inner diameter is equivalent to oocyte diameter 1 / 5 ~ 1 / 4) holding oviposit tubes on the needle pulling instrument and forging instrument. Debug the micro-operating system.
[0024] Human oocytes (MII) to be enucleated from which granulosa cells are to be removed are transferred to HTF-HEPES micro-droplets and placed on a micromanipulator inverted microscope. see image 3 , hold the oocyte with a wide inner diameter tube, and place the polar body at the 6 o'clock or 12 o'clock position.
[0025] see Figure 4 , the assisted hatching needle cuts the zona pellucida at the first polar body, and...
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The invention provides a method for removing a mammal oocyte nucleus through using a wide-internal-diameter oocyte holding tube. The method adopts the wide-internal-diameter oocyte holding tube with the internal diameter being 1 / 5-1 / 4 of the diameter of an oocyte to hold the oocyte, an auxiliary hatching needle cuts a clear tape at the position of a first polar body, the wide-internal-diameter oocyte holding tube horizontally aims at the crack of the clear tape and is used to apply a negative pressure to suck out the first polar body and 1 / 10-1 / 8 of a cytoplasm, and the auxiliary hatching needle is used to shift or rotate the oocyte in order to separate the sucked-out nucleus carried cytoplasm from the oocyte. In the invention, the oocyte holding tube simultaneously has oocyte holding and nucleus suck-out effects, so the method has the advantages of simple operation, reduction of a nucleus removal step, shortening of the nucleus removal operation time, nucleus removal success rate improvement, reduction of the damages to the oocyte, and benefiting for the growth of an embryo in the subsequent nucleus transplantation process.
Description
technical field [0001] The invention relates to a method for removing oocyte nuclei in the technical field of bioengineering, in particular to a method for removing mammalian oocyte nuclei with a wide inner diameter ovipositor tube. Background technique [0002] Mammalian somatic cell nuclear transfer technology refers to the use of oocytes with denuclearized oocytes as recipients and somatic cell nuclei as donors to fuse the two to construct reconstructed embryos by artificial means, such as transplanting the embryos into the uterus of a surrogate mother and producing Individuals are called cloned animals; if reconstituted embryos are used to establish embryonic stem cell lines, they are called somatic cell nuclear transfer embryonic stem cells. [0003] Mammals such as sheep, cattle, pigs, mice, and rabbits and various nuclear transfer embryonic stem cell lines have been born through somatic cell nuclear transfer technology. This technology has opened up broad application...
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Inventor 谌兵来高擎曾桥
Owner ZHEJIANG CELLPRO BIOTECH