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Leech cell in-vitro culture medium and culture method thereof

A culture method and medium formula technology are applied in the field of in vitro culture of leech cells to achieve the effect of accurate and reliable experimental basis

Active Publication Date: 2018-07-10
长握生物科技(江苏)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the in vitro culture technology of leech cells is still in its infancy, and the selection of its medium, the determination of culture conditions, the selection of antibiotics, etc. are all problems that need to be overcome urgently.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A kind of leech living body growth cycle selection method provided by the present invention, concrete steps are as follows

[0043] Step 1: Soak 1-year-old, 2-year-old, and 3-year-old living leeches in normal saline for 1 hour to initially remove impurities in the body and ensure the vitality of the somatic cells to the maximum extent, then wash the surface mucous membrane with normal saline until it is clean, and finally wash it three times with distilled water Then soak in 70% alcohol for 5 minutes to remove bacteria on the body surface, and rinse with distilled water several times to remove alcohol;

[0044] Step 2: Transfer the leeches after the skin disinfection treatment in step 1 to the ultra-clean workbench, put them into a sterile petri dish filled with sterile PBS, and cut off the leeches with sterile scissors and tweezers Take the head tissue from both ends of the body;

[0045] Step 3: transfer the head tissue in step 2 to another sterile petri dish equippe...

Embodiment 2

[0068] A kind of leech cell primary culture optimized insect cell culture medium provided by the present invention, the specific steps are as follows

[0069] Step 1: Soak the living leeches in normal saline for 1 hour to initially remove impurities in the body and ensure the vitality of the somatic cells to the maximum extent, then wash the surface mucous membranes with normal saline until clean, and finally wash them three times with distilled water and place them in 70% alcohol Soak in water for 5 minutes to remove bacteria on the body surface, and rinse with distilled water several times to remove alcohol;

[0070] Step 2: Transfer the leech after the skin disinfection treatment in step 1 to the ultra-clean workbench, and put it into a sterile petri dish filled with 600U / mL of PBS of different antibiotics, use sterile scissors and The tweezers cut off both ends of the leech body to get the head tissue;

[0071] Step 3: transfer the head tissue in step 2 to another sterile...

Embodiment 3

[0092] Antibiotic class selection provided by the present invention, concrete steps are as follows

[0093] Step 1: Soak the living leeches in normal saline for 1 hour to initially remove impurities in the body and ensure the vitality of the somatic cells to the maximum extent, then wash the surface mucous membranes with normal saline until clean, and finally wash them three times with distilled water and place them in 70% alcohol Soak in water for 5 minutes to remove bacteria on the body surface, and rinse with distilled water several times to remove alcohol;

[0094] Step 2: Transfer the leech after the skin disinfection treatment in step 1 to the ultra-clean workbench, and put it into the aseptic culture of PBS containing different kinds of antibiotics (penicillin, streptomycin and amphotericin B) In the dish, use sterile scissors and tweezers to cut off the two ends of the verruca to take the head tissue;

[0095] Step 3: Transfer the head tissue from step 2 to another st...

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PUM

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Abstract

The invention discloses a leech cell in-vitro culture medium and a culture method thereof. According to the method, sheared leech tissue blocks are inoculated into a cell sealed culture bottle; an Insect-CCulture insect cell culture medium is added; still standing culture is performed in a cell culture box; then, a tissue block recovery method is matched for separation culture; finally, purified leech cells are obtained through a trypsin digestion method for further culture. Compared with other insect cell culture media, the method has the advantages that the obtaining of the leech purified body cells can be successfully obtained at a higher speed; the preparation is made for further performing the leech cell in-vitro subculture; in addition, a set of in-vitro primary culture leech cell system is built.

Description

technical field [0001] The invention relates to a method for culturing leech cells in vitro, belonging to the field of bioengineering. Background technique [0002] With the rapid development of life sciences, cell culture is an essential process for both the entire bioengineering technology and one of the biological cloning technologies, and promotes the development of cell biology, molecular biology, biochemistry development in other fields. [0003] Leeches (Whitmania pigra Whitman) belong to the family Annelidae, Hirudoidae. According to statistics, there are more than 600 species of leeches distributed all over the world, and nearly 100 species are distributed in my country. Leeches are cold-blooded annelids that can grow and reproduce in both the north and the south of China. They mainly live in freshwater reservoirs, ditches, paddy fields, lakes and swamps, and most often in ponds rich in organic matter or in small rivers without pollution. The optimum temperature f...

Claims

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Application Information

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IPC IPC(8): C12N5/07
CPCC12N5/0601C12N2500/12C12N2500/16C12N2500/30C12N2500/32C12N2500/34C12N2500/38
Inventor 虞龙
Owner 长握生物科技(江苏)有限公司
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