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Vitro culturing method of pearl shell mantle tissue and culture medium

A technology of in vitro culture and mantle, applied in tissue culture, biochemical equipment and methods, fish farming, etc., can solve the problems affecting the structure change of in vitro pearl sacs, high mortality rate of pearl oysters, and low pearl formation rate , to achieve high survival rate and beading rate, reduce pollution opportunities, and improve survival rate

Active Publication Date: 2012-10-10
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 filtering and sterilizing the liquid in the body of Pinctada martensii, and shellfish serum is obtained from the heart or large blood vessels of pearl oysters, after Centrifuge, get supernatant to obtain pearl oyster serum), add 0.1-5% hydrolyzed pearl liquid 10ml, and add 100IU / ml penicillin 1ml, 0.1g / L streptomycin 1ml, adjust the pH value to 6.8-7.2. The preparation of hydrolyzed pearl liquid is to mix nacre powder and honey with pH 2.5-6 at a ratio of 0.1-5:100, then stir at room temperature above 5°C, or stir at room temperature and then heat and stir step by step with water for 2-3 hours. 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 filtering and sterilizing the liquid in the body of Pinctada martensii, and shellfish serum is obtained from the heart or large blood vessels of pearl oysters, after Centrifuge, get the supernatant to obtain pearl oyster serum), add 0.1-5% hydrolyzed pearl liquid 10ml, and add 100IU / ml ampicillin 2ml, or 0.1g / L streptomycin sulfate 2ml, adjust the pH value to 6.8-7.2. The preparation of hydrolyzed pearl liquid is to soak the cleaned mother-of-pearl in acetic acid, soften it, and grind it mechanically to obtain pure pearl flakes. And, vacuum concentration, activated carbon decolorization and impurity removal, to obtain pearl stock solution.

[0046] Two, the in vitro culture method of pearl oyster mantle tissue:

[0047] Example 1

[0048] (1) Take trypsin in proportion, dissolve it in three-distilled water to prepare a 0.1-0.5% trypsin...

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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 1mm3 to 2mm3 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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Patent Type & Authority Patents(China)
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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