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Vitro culturing method of pearl shell mantle tissue and culture medium

An in vitro culture, mantle technology, applied in tissue culture, biochemical equipment and methods, fish farming, etc., can solve the problems of in vitro pearl sac structure changes, high mortality of pearl oysters, low pearl formation rate, etc. , to achieve the effect of high survival rate and beading rate, reducing the chance of pollution and improving the survival rate

Active Publication Date: 2009-07-29
广西中医学院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, when the in vitro pearl sac formed by the mantle tissue block is used in the experiment of inserting the pearl, the pearl-forming rate is low, the number of pearls is high, and the mortality rate of the pearl oyster is high. This is all related to the inserting technique. The size and the way the bead nucleus is transported to the predetermined nucleus through the incision directly affect the structural changes of the pearl sac in vitro

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Take 100ml of boiled natural seawater, add 2-10ml of shellfish tissue fluid or shellfish serum, (shellfish tissue fluid is obtained by filtration and sterilization of the liquid in Pinctada martensii, and shellfish serum is taken from the heart or large blood vessels of pearl shellfish. Centrifuge, take the supernatant to obtain pearl oyster serum), add 10ml of 0.1-5% hydrolyzed pearl liquid, and add 1ml of 100IU / ml penicillin, 1ml of 0.1g / L streptomycin, and adjust the pH value to 6.8-7.2. The preparation of the hydrolyzed pearl liquid is to mix nacre powder and honey with a pH of 2.5 to 6 at a ratio of 0.1 to 5:100, and then stir at room temperature above 5 °C, or stir at room temperature and then heat and stir for 2 to 3 hours in sections through water. get.

Embodiment 2

[0045] Take 100ml of boiled natural seawater, add 2-10ml of shellfish tissue fluid or shellfish serum, (shellfish tissue fluid is obtained by filtration and sterilization of the liquid in Pinctada martensii, and shellfish serum is taken from the heart or large blood vessels of pearl shellfish. Centrifuge, take the supernatant to obtain pearl oyster serum), add 10ml of 0.1-5% hydrolyzed pearl liquid, and add 2ml of 100IU / ml ampicillin, or 2ml of 0.1g / L streptomycin sulfate, and adjust the pH value to 6.8-7.2. The preparation of the hydrolyzed pearl liquid is to soak the washed mother-of-pearl with acetic acid, soften it, and grind it mechanically to obtain a pure nacre sheet, which is dried, pulverized, ground, hydrolyzed by heating with sulfuric acid, and dissolved in calcium carbonate or lime milk. And, vacuum concentration, activated carbon decolorization and impurity removal to obtain pearl stock solution.

[0046] 2. In vitro culture method of pearl oyster mantle tissue: ...

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Abstract

The invention discloses an in-vitro culture method for the pearl shell mantle tissue and a special culture solution thereof. The culture medium comprises hydrolyzed pearl liquor, shell liquor, antibiotics and seawater. The method conventionally comprises the following steps: taking the material from a nacre to obtain the mantle epithelium, removing the skirt band part, cutting the tissue into tissue blocks with the size of about 1mm to 2mm after purification treatment, putting the tissue blocks in the protease solution for digestion, and adding the culture solution to stop the digestion. The preliminarily digested tissue blocks are respectively put into a culture dish or cling to the pearl core which is covered by adherence substance. The cultured pearl shell mantle tissue grows in good conditions; after 4 hours, the cells are emigrated from the tissue blocks; and after 2 days in-vitro pearl sacs are formed effectively. The method can be used to inject the anucleate pearl sacs at different positions in the pearl shell with the inserted pearl nucleus so as to culture the anucleate pearl. The method has low cost, a high survival rate, a high pearl formation rate, and little damage to the nacre.

Description

technical field [0001] The invention relates to a tissue culture technology and application field of aquatic invertebrates, in particular to an in vitro culture method and culture medium of pearl oyster mantle tissue. Background technique [0002] Pearls are formed by the secretion of nacre from the pearl sac in the pearl oyster. The pearl sac is formed by the proliferation of the epithelial cells of the mantle of the pearl oyster after being stimulated. Therefore, the mantle of the pearl oyster plays an important role in the formation of pearls. The modern artificial pearl culture method is to implant the mantle tissue block of the donor pearl oyster alone or together with the bead nucleus into the recipient pearl oyster to form a seedless pearl or a nucleated pearl. During the implantation operation, the position and tightness of the epithelial cells attached to the bead nucleus, the area and shape of the tissue block, the location of the material, the position of the nucl...

Claims

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Application Information

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IPC IPC(8): A01K61/00C12N5/06
CPCY02A40/81
Inventor 林江林湧邓家刚王乃平甘霖王勤单华林海林其溪杨辉戴铭李中华刘强陈明伟杨继峰
Owner 广西中医学院
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