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Gelatin microsphere and preparation method thereof

A technology of gelatin and microspheres, applied in the field of cell culture, can solve the problems of unfavorable cell expansion, cytotoxic side effects, etc., and achieve the effects of improving cell activity, good degradation effect, and reducing toxicity

Inactive Publication Date: 2017-08-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, commonly used aldehyde cross-linking agents (such as glutaraldehyde) have certain toxic side effects on cells, which is not conducive to cell expansion.

Method used

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  • Gelatin microsphere and preparation method thereof
  • Gelatin microsphere and preparation method thereof
  • Gelatin microsphere and preparation method thereof

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preparation example Construction

[0028] The invention provides a kind of preparation method of gelatin microsphere, comprises the following steps:

[0029] First, dissolve the gelatin in water. The dosage of the gelatin is preferably 10%-20% w / v. Dissolve in water and heat to dissolve. The heating is preferably 70°C-100°C. Add the oil phase in which emulsifier A is dissolved and stir. The emulsifier A is preferably any one or more of liquid paraffin, castor oil, olive oil, peanut oil, soybean oil, cocoa butter, coconut oil vinegar, palm oil vinegar, ethyl acetate, beeswax and silicone oil, More preferably, it is any one or more of liquid paraffin, ethyl acetate, olive oil, peanut oil, and soybean oil. The volume ratio of gelatin volume to emulsifier A is preferably 1-5:1. Stir at a certain speed, preferably 100r / min-900r / min, and then add emulsifier B for stirring. The emulsifier B is preferably any one or more of liquid paraffin, castor oil, olive oil, peanut oil, soybean oil, cocoa butter, coconut oil ...

Embodiment 1

[0036] Embodiment 1 prepares gelatin microsphere

[0037] (1) Weigh 5g of gelatin, dissolve it in 50mL of ultrapure water, and heat it at 70°C to dissolve completely. The gelatin solution is stirred into the oily phase in which the emulsifier liquid paraffin is dissolved. The volume ratio of gelatin volume to emulsifier liquid paraffin is 5:1, stirring at a speed of 900r / min to form an emulsion. The emulsifier olive oil is then added to form a second oily phase. The volume ratio of the gelatin volume to the emulsifier olive oil is 1:2, and then stirred at a speed of 450rpm for 1min.

[0038] (2) Cool the emulsion in an ice-water bath to below 5 degrees Celsius, and continue to stir at a speed of 450 rpm for 30 minutes, then add 2 times the volume of the emulsion in an ice-bathed dehydration and deoiling agent absolute ethanol at -20°C, and stir dehydration. The gelatin microspheres were washed alternately with dehydrating and deoiling reagents isopropanol and acetone, and ...

Embodiment 2

[0040] Embodiment 2 prepares gelatin microsphere

[0041] (1) Weigh 5g of gelatin, dissolve it in 50mL of ultrapure water, and heat it at 70°C to dissolve completely. The gelatin solution is added with stirring to the oil phase in which the emulsifier ethyl acetate is dissolved. The volume ratio of gelatin volume to emulsifier ethyl acetate is 2:1, stirring at a speed of 900r / min to form an emulsion. Then add the emulsifier soybean oil to form the second oil phase. The volume ratio of gelatin to emulsifier soybean oil is 1:3. Then stir for 1 min at a speed of 450 rpm.

[0042] (2) Then rapidly cool the emulsion to below 5 degrees Celsius in an ice-water bath, and continue to stir at a speed of 450 rpm for 30 minutes. Then add 2 times the volume of the emulsion in a -20°C ice-bath dehydration and deoiling agent isopropanol, and stir for dehydration . The gelatin microspheres were washed alternately with dehydrating and deoiling reagents isopropanol and acetone, and finally...

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Abstract

The invention relates to the field of cell culture, in particular to a gelatin microsphere and a preparation method thereof. The preparation method of the gelatin microsphere provided by the invention comprises the steps that gelatin is dissolved into water; different emulsifying agents are respectively added; stirring is performed to obtain emulsion; different dehydrating agents are added into the emulsion; cleaning, drying and sieving are performed; then, gelatin microsphere crude products are obtained; an ethanol solution containing Jinniping crosslinked curing agents is added into the gelatin microsphere crude products; the gelatin microsphere is obtained through preparation. Experimental results show that the gelatin microsphere prepared by the preparation method provided by the invention has smaller toxic action on cells; the toxicity on the cells can be reduced in the cell amplification culture process; the cell activity is improved and the degradation effect is better.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a gelatin microsphere and a preparation method thereof. Background technique [0002] Gelatin is a polymer material of natural origin and good biocompatibility, which is a product obtained by mild and irreversible degradation of collagen. Gelatin contains keratin, elastin, melanin and chondroitin, which can promote cell adhesion and growth, which is an important factor for gelatin to be used as a microcarrier for cell expansion. When the temperature drops to about 40°C, gelatin will undergo a transformation from a free coil to a helical conformation. Therefore, gelatin cannot be used directly under physiological conditions. If it is to be used as a cell expansion carrier, it must be properly cross-linked. [0003] At present, commonly used aldehyde cross-linking agents (such as glutaraldehyde) have certain toxic side effects on cells, which is not conducive to cell expansion. Conte...

Claims

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Application Information

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IPC IPC(8): C08J3/24C08J3/05C08L89/00C08K5/1545
CPCC08J3/24C08J3/05C08J2389/00C08K5/1545C08L89/00
Inventor 陈海佳葛啸虎王一飞戚康艺张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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