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Acipenser dabryanus testis cell cryopreservation liquid as well as testis cell cryopreservation method and application

A cryopreservation solution and cryopreservation technology, which can be used in applications, preservation of human or animal bodies, animal husbandry, etc., can solve the problems of cryopreservation of testicular cells of Acipenser dabryi which have not yet been seen, and achieve short equilibrium time, high survival rate and The effect of proliferation rate

Active Publication Date: 2016-02-03
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There is no report on the cryopreservation of testicular cells of Acipenser dabryi

Method used

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  • Acipenser dabryanus testis cell cryopreservation liquid as well as testis cell cryopreservation method and application
  • Acipenser dabryanus testis cell cryopreservation liquid as well as testis cell cryopreservation method and application
  • Acipenser dabryanus testis cell cryopreservation liquid as well as testis cell cryopreservation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A frozen dilution of Acipenser dabryi, comprising:

[0051]

[0052]

[0053] The rest is L-15 culture medium, and the pH of the preservation solution is 8.0.

[0054] The pH of the above preservation solution was passed through 1M NaHCO 3 or 25mM HEPES adjustment.

[0055] The preparation method of described Dabry's sturgeon frozen dilution comprises the following steps:

[0056] (1) Use a pipette to accurately measure the volume of each liquid component required to prepare the diluent:

[0057] 1) Measure 3ml of L-15 culture solution into a centrifuge tube.

[0058] 2) Take 1ml of fetal bovine serum and 500μl of ethylene glycol, and add them to the L-15 culture medium.

[0059] (2) Accurately weigh the weight of the solid components required for the diluent, with an accuracy of 0.1 mg: bovine serum albumin 250.0 mg, trehalose 94.6 mg, lecithin 400.0 mg, and dissolve in the diluent.

[0060] (3) Finally add L-15 culture medium to make the volume to 5ml. Ste...

Embodiment 2-5

[0062] A frozen dilution of Acipenser dabryi, comprising:

[0063]

[0064] The pH of the above preservation solution was passed through 1M NaHCO 3 or 25mM HEPES adjustment.

Embodiment 7

[0066]A method for cryopreservation of testicular cells of Dabry's sturgeon, comprising the following steps:

[0067] 5ml of Acipenser dabryi testis cell suspension (5×10 5 Dabry's Acipenser testis cells) as an example to illustrate the method of cryopreservation of Acipenser dabryi's testis cells. The formula of the cryopreservation solution used in this embodiment is as shown in Example 1. The following operations were all carried out in a sterile biological safety cabinet.

[0068] Preparation of Dabry's testicular cell suspension:

[0069] 1) Move the cleaned testis tissue to a petri dish, and cut the testis tissue to 1mm 3 small pieces. Add 0.25% trypsin digestion solution at the ratio of digestion solution: tissue block = 4ml: 1g (v / w), digest in a 20°C incubator, and pipette once every 30min.

[0070] 2) After digesting the testis tissue in step 1) for 3 hours, add L-15 medium + 20% fetal bovine serum to terminate the digestion, and filter through a 40 μm filter. ...

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PUM

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Abstract

The invention provides acipenser dabryanus testis cell cryopreservation liquid as well as a testis cell cryopreservation method and application. The cryopreservation liquid comprises bovine serum albumin, trehalose, ethylene glycol, lecithin and fetal calf serum. Meanwhile, the invention further provides an acipenser dabryanus testis cell cryopreservation method. By using the method, damages of ice crystals on cells can be reduced to the greatest extent by using a relatively low cooling manner under the condition that liquid-nitrogen cooling is not used, and the cells have high survival rate and reproduction rate after recovery, so that warm cryopreservation is possible to realize, and long-period stable cell supply is provided for transplanting acipenser dabryanus spermatogonia.

Description

technical field [0001] The invention relates to the technical field of cell preservation of Acipenser dabryi, in particular to a cryopreservation liquid for testicular cells of Acipenser dabryi and a method and application thereof. Background technique [0002] In the male gonad, spermatozoa originate from spermatogonia stem cells (SSCs), which are capable of both self-replication and differentiation into mature spermatozoa. Mammalian spermatozoa are produced in the seminiferous tubules. In murines, it has been confirmed functionally that some type A spermatogonia exist in the basement membrane of the vas deferens, and the evaluation of stemness has been completed through chemotherapy, radiotherapy and transplantation techniques. In the testes of fish, spermatozoa occur in seminal vesicles. Germ cells reside within the cytoplasmic extensions of Sertoli cells forming the seminal vesicles, and each seminal vesicle germ cells proliferate and differentiate simultaneously. Thi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 颉璇厉萍危起伟席萌丹郭威乔新美杜浩刘志刚
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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