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Prostate extracellular matrix gel for cultivating prostate cells

A cell culture and prostate technology, applied in the field of prostate extracellular matrix gel, can solve the problems of inconsistent physiological state of cell growth, difficulty in simulating the natural growth environment of cells, etc., and achieve good biocompatibility and prolongation of proliferation period

Inactive Publication Date: 2016-02-10
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Using the existing technology to culture cells, the cells show a planar growth mode. The cells grow on the surface of the medium and only extend along the plane of the medium. This is not consistent with the physiological state of cell growth in mammals, and it is difficult to simulate the growth of cells in lactation. The natural growth environment in the animal body, which is contrary to the original intention of human cell culture

Method used

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  • Prostate extracellular matrix gel for cultivating prostate cells
  • Prostate extracellular matrix gel for cultivating prostate cells
  • Prostate extracellular matrix gel for cultivating prostate cells

Examples

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Embodiment 1

[0029] Prostate extracellular matrix gel for prostate cell culture, the preparation method of the prostate extracellular matrix gel comprises the following steps:

[0030]A. Preparation of prostate extracellular matrix: take a male adult mammal bladder tissue section under sterile conditions, the size of the section is 1cm×1cm×0.5cm; take 10 sections, add 5mL of aprotinin solution with a concentration of 11kIU / mL , stirred at 4°C for 55h; then stirred in 5mL of 1.2% polyethylene glycol octyl phenyl ether solution at 4°C for 20h; then in 10mL of 0.75% dodecyl Stir in sodium sulfate solution at 4°C for 30h; then stir in 5mL of DNA hydrolase at 45U / mL and RNase A at 1.5U / mL for 20h at 4°C; buffer with 1×PBS The solution was washed three times, each time for 5min, refrigerated at 4°C, then freeze-dried under vacuum and passed through 10-15kGyCo 60 Sterilized by irradiation and stored at -20°C; the freeze-drying temperature under vacuum conditions is -10°C, and the vacuum degree i...

Embodiment 2

[0034] Prostate extracellular matrix gel for prostate cell culture, the preparation method of the prostate extracellular matrix gel comprises the following steps:

[0035] A. Preparation of prostate extracellular matrix: Take a male adult mammalian bladder tissue slice under sterile conditions, the size of the slice is 1cm×1cm×0.5cm; take 10 slices, add 15mL of aprotinin solution with a concentration of 9kIU / mL , stirred at 4°C for 45h; then stirred in 10mL of 0.85% polyethylene glycol octyl phenyl ether solution at 4°C for 30h; then in 5mL of 1.5% dodecyl Stir in sodium sulfate solution at 4°C for 20h; then stir in 15mL of DNA hydrolase at a concentration of 55U / mL and RNase A at a concentration of 0.5U / mL at 4°C for 28h; buffer with 1×PBS The solution was washed three times, each time for 5min, refrigerated at 4°C, then freeze-dried under vacuum and passed through 10-15kGyCo 60 Sterilized by irradiation and stored at -20°C; the freeze-drying temperature under the vacuum con...

Embodiment 3

[0039] Prostate extracellular matrix gel for prostate cell culture, the preparation method of the prostate extracellular matrix gel comprises the following steps:

[0040] A. Preparation of prostate extracellular matrix: take a male adult mammalian bladder tissue slice under sterile conditions, the size of the slice is 1cm×1cm×0.5cm; take 10 slices, add 8mL of aprotinin solution with a concentration of 10kIU / mL , stirred at 4°C for 48h; then stirred in 5mL of 1% polyethylene glycol octylphenyl ether solution at 4°C for 24h; then in 7.5mL of 1% dodecane Stir at 4°C for 24h in sodium bisulphate solution; then stir at 4°C for 24h in 10mL of DNA hydrolase concentration of 50U / mL and RNase A at 1U / mL; buffer with 1×PBS The solution was washed three times, each time for 5 minutes, refrigerated at 4°C, and then freeze-dried under the vacuum condition of the vacuum bar at -15°C, and the vacuum degree was 11Pa;

[0041] B. Preparation of prostate extracellular matrix gel: freeze the s...

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Abstract

The invention relates to prostate extracellular substrate gel for cultivating prostate cells. The prostate extracellular matrix gel has the advantages that the prostate extracellular matrix gel is white and semi-transparent and is excellent in biocompatibility; the human prostate epithelial cells can be cultivated by the aid of the prostate extracellular matrix gel and are in tufted three-dimensional spatial growth modes which are consistent with in-vivo cell growth physiological status of mammals, and accordingly in-vivo natural growth environments for the cells in the mammals can be effectively simulated; proliferative stages are long (11 days) and are prolonged by 6 days as compared with proliferative stages of cells cultivated in the prior art.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a prostate extracellular matrix gel for prostate cell culture. Background technique [0002] Cell culture technology, also called cell cloning technology, is the core and basic technology in the field of biotechnology. The general process of cell culture includes 1) Preparation: cleaning, drying, disinfection of utensils, preparation of medium and other reagents, packaging and sterilization, cleaning and disinfection of sterile rooms or ultra-clean benches, incubators and other instruments 2) Obtaining materials: take out tissue cells from the body in a sterile environment, and insert them into culture vessels after treatment; 3) Culture: the process of inserting the obtained tissue cells into culture flasks or culture plates is called culture . The cells in culture should be observed every once in a while. The observations include whether the cells grow well, whether the shape is n...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 陈伟
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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