Method for extracting mesenchymal stem cells from umbilical cord Wharton's jelly

A technology for Wharton's jelly and stem cells, applied in the field of bioengineering, can solve the problems of insufficient adherence and growth support ability of primary cells, and achieve the effects of rapid preparation and identification, high yield, and shortening of separation and acquisition time.

Inactive Publication Date: 2015-07-08
HUBEI SHENGMINGYUAN STEM CELL CO LTD
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the common problem of the current serum-free medium is that it has ins

Method used

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  • Method for extracting mesenchymal stem cells from umbilical cord Wharton's jelly
  • Method for extracting mesenchymal stem cells from umbilical cord Wharton's jelly
  • Method for extracting mesenchymal stem cells from umbilical cord Wharton's jelly

Examples

Experimental program
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Embodiment

[0029] 1. Obtain 10 cm of umbilical cord tissue of a caesarean section newborn, and fully remove blood stains with 3 wt % PBS buffer solution containing aminopenicillin and streptomycin sulfate.

[0030] 2. Cut the washed umbilical cord evenly into 3-4cm long sections, perform mechanical separation, bluntly peel off Wharton's jelly, and remove the umbilical artery and umbilical vein at the same time.

[0031] 3. Cut the peeled Wharton's glue into 1mm 3 -3mm 3 Add 10 mL of collagenase A with a concentration of 3 mg / mL, place at 37 °C, 5% CO 2 The digestion process was carried out in the incubator for 2 hours, during which it was taken out every 20 minutes to detect the digestion situation.

[0032] 4. After the digestion process, centrifuge to collect the cells and residual tissue, and use 2wt% penicillin-streptomycin and no Ca 2+ , Mg 2+ Wash with PBS buffer to remove residual collagenase A, centrifuge at 2000 rpm for 5 minutes, and discard the supernatant.

[0033] 5. Sl...

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Abstract

The invention belongs to the technical field of bioengineering, and in particular relates to a method for extracting mesenchymal stem cells from umbilical cord Wharton's jelly. The method comprises the steps of separating the Wharton's jelly, digesting the Wharton's jelly, centrifugally separating liquid after being digested, processing separated liquid at low temperature, centrifugally separating liquid after being processed at low temperature, culturing and the like. The method for extracting the mesenchymal stem cells from the umbilical cord Wharton's jelly provided by the invention is a method for obtaining mesenchymal stem cells of an umbilical cord with high efficiency and high purity; the yield and the purity of the mesenchymal stem cells of the umbilical cord are increased by utilizing enzymic digestion in combination with gelatin spreading; the separation and obtaining time is effectively shortened; a method for rapidly preparing and identifying is realized; and the yield, which is higher than the yield in the previous culture method, can be obtained.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for extracting mesenchymal stem cells from umbilical cord Wharton's jelly. Background technique [0002] The umbilical cord of a normal person contains three blood vessels: two umbilical arteries (PluralArteriae umbilicales) and one umbilical vein (Vena umbilicalis). Two arteries carry carbon dioxide-rich and nutrient-poor blood to the placenta, and veins carry oxygen-rich and nutrient-rich blood back to the fetus. Another artery degenerated during the first week of the fetus. In addition to the pulmonary artery / vein, there are only umbilical arteries / veins in which arteries transport venous blood (blood rich in carbon dioxide and waste) and veins transport arterial blood (blood rich in oxygen and nutrients). The human umbilical cord is about 50 to 60 cm long and about 2 cm in diameter. The blood vessels in the umbilical cord are wrapped by Wharton's...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 王锐峰
Owner HUBEI SHENGMINGYUAN STEM CELL CO LTD
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