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Digestive enzyme composition and application thereof, and isolated culture method of umbilical epithelial cells

A technology of separation culture and cell culture, applied in animal cells, vertebrate cells, biochemical equipment and methods, etc., can solve the problems of low cell purity, low total number of cells, long culture time, etc., to promote cell proliferation, increase cell Total, reduced time effects

Inactive Publication Date: 2015-09-09
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0023] However, the above-mentioned existing method for obtaining tissue epithelial cells takes a long time for primary culture, the total number of cells obtained is low, and the purity of cells is low. Doping with fibroblasts

Method used

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  • Digestive enzyme composition and application thereof, and isolated culture method of umbilical epithelial cells
  • Digestive enzyme composition and application thereof, and isolated culture method of umbilical epithelial cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Primary isolation and culture of embodiment 1 human umbilical cord epithelial cells

[0070] (1) Gelatin coating of Petri dishes

[0071] Weigh gelatin powder (Sigma), make a 0.1% (W / V) solution with deionized water, dissolve in a water bath at 37°C, filter it through a 0.22μm filter while it is hot, and place it at 4°C for later use.

[0072] Take the prepared 0.1% gelatin solution for rewarming, add 0.3mL / cm 2 0.1% gelatin solution, spread evenly on the bottom of the dish, place it in a 37°C cell culture incubator for 24 hours, then suck out all the gelatin in the dish and discard it, and the gelatin coating of the cell dish is completed.

[0073] (2) Primary isolation of human umbilical cord epithelial cells

[0074] Umbilical cords were collected after postpartum disposal by pregnant women in the hospital. Take an umbilical cord under sterile conditions, wash the blood on the surface of the umbilical cord with 100 mL of sterile phosphate buffered saline (PBS), th...

Embodiment 2

[0079] Example 2 Primary isolation and culture of human umbilical cord epithelial cells

[0080] (1) Gelatin coating of Petri dishes

[0081] Weigh gelatin powder (Sigma), make a 0.1% (W / V) solution with deionized water, dissolve in a water bath at 37°C, filter it through a 0.22μm filter while it is hot, and place it at 4°C for later use.

[0082] Take the prepared 0.1% gelatin solution for rewarming, add 0.3mL / cm 2 0.1% gelatin solution, spread evenly on the bottom of the dish, place it in a 37°C cell culture incubator for 24 hours, then suck out all the gelatin in the dish and discard it, and the gelatin coating of the cell dish is completed.

[0083] (2) Primary isolation of human umbilical cord epithelial cells

[0084] Umbilical cords were collected after postpartum disposal by pregnant women in the hospital. Take an umbilical cord under sterile conditions, wash the blood on the surface of the umbilical cord with 100 mL of sterile phosphate buffered saline (PBS), then ...

Embodiment 3

[0089] Example 3 The Primary Isolation and Culture of Human Umbilical Cord Epithelial Cells

[0090] (1) Gelatin coating of Petri dishes

[0091] Weigh gelatin powder (Sigma), make a 0.1% (W / V) solution with deionized water, dissolve in a water bath at 37°C, filter it through a 0.22μm filter while it is hot, and place it at 4°C for later use.

[0092] Take the prepared 0.1% gelatin solution for rewarming, add 0.3mL / cm 2 0.1% gelatin solution, spread evenly on the bottom of the dish, place it in a 37°C cell culture incubator for 24 hours, then suck out all the gelatin in the dish and discard it, and the gelatin coating of the cell dish is completed.

[0093] (2) Primary isolation of human umbilical cord epithelial cells

[0094] Umbilical cords were collected after postpartum disposal by pregnant women in the hospital. Take an umbilical cord under sterile conditions, wash the blood on the surface of the umbilical cord with 100 mL of sterile phosphate buffered saline (PBS), the...

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Abstract

The invention relates to the technical field of cell isolated culture, particularly a digestive enzyme composition and application thereof, and an isolated culture method of umbilical epithelial cells. The digestive enzyme composition comprises hyaluronidase, collagenase VIII and pancreatin. The umbilical epithelial cell primary isolation adopts the hyaluronidase, collagenase VIII and pancreatin for combined digestion. The digestive enzyme composition has the advantages of mild properties, low cell trauma and high enzymolysis rate and greatly enhances the epithelial cell isolation efficiency.

Description

technical field [0001] The invention relates to the technical field of cell separation and culture, in particular to a digestive enzyme composition and application thereof, and a method for separation and culture of umbilical cord epithelial cells. Background technique [0002] The umbilical cord is a tubular structure that connects the fetus to the placenta. It was originally formed by the amniotic membrane surrounding the stalk-shaped extension of the yolk sac and urine membrane. The arteries and veins of the umbilical cord are formed through the blood vessels of the urethra, and the blood vessels of the yolk sac form the umbilical mesenteric artery and the umbilical mesenteric vein. When the yolk sac and its blood vessels degenerate, the umbilical artery and umbilical vein develop, and loose gelatinous mesenchyme can be seen in these spaces. The umbilical cord carries CO between the blood of the mother and the fetus 2 and O 2 , the place where metabolic waste and nutri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N9/50C12N9/94C12N5/071
CPCC12Y302/01035C12N5/0605C12N9/6489C12N9/94C12Y304/24
Inventor 陈海佳王一飞葛啸虎吴子杰王小燕李平
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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