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Method for obtaining MSCs (mesenchymal stem cells) from umbilical cord

A technology of stem cells and umbilical cord, applied in the field of cell preparation, can solve the problems of increasing safety hazards and long operation time, and achieve the effect of preventing damage

Inactive Publication Date: 2019-03-15
厦门博瑞思生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method makes it necessary to add antibiotics to prevent contamination in subsequent cultures, which increases the safety hazards of clinical application.
Simultaneous operation time is long

Method used

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  • Method for obtaining MSCs (mesenchymal stem cells) from umbilical cord

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Screening of puerpera: screening of infectious disease test results, family history and genetic history of puerpera, selection of qualified infectious disease test (including but not limited to hepatitis B, hepatitis C, syphilis, AIDS, cytomegalovirus), no family history and genetic history For full-term expectant mothers who have caesarean section, after the fetus is delivered, it is confirmed that the fetus is healthy and has no deformities or congenital diseases.

[0032] Umbilical cord collection and transportation: After the fetus is delivered, the umbilical cord is cut off routinely, and about 10mL of cord blood is collected into a blood collection tube for subsequent testing. The middle 10-20cm umbilical cord is selected, cleaned and disinfected with alcohol gauze, cut off, and put into the umbilical cord for storage and transportation. In the bottle, 10 mL of DMEM / F12 medium containing 100 U / mL penicillin and streptomycin was added to the storage bottle in advanc...

Embodiment 2

[0041] Screening of puerpera: same method and requirements as in Example 1.

[0042] Collection of umbilical cord and transportation: the same method and requirements as in Example 1.

[0043] Washing and disinfection of the umbilical cord: with the method and requirements of Example 1, only Aner Iodine disinfectant is used.

[0044] Remove blood vessels and separate Wharton's jelly: Transfer the cleaned umbilical cord from the centrifuge tube to a petri dish, cut off 1 cm from the head and tail and discard, and cut the rest into 2 cm sections. Take a section of umbilical cord, peel off the outer skin with toothed forceps, remove the veins and arteries, and peel off the Wharton's jelly, taking care not to peel off the epidermal tissue of the umbilical cord. Wharton's jelly was placed in a 50mL centrifuge tube and washed twice with cleaning solution.

[0045] Transfer Wharton's jelly to a new centrifuge tube, add serum-free medium to make it just submerge Wharton's jelly, use...

Embodiment 3

[0051] Screening of puerpera: same method and requirements as in Example 1.

[0052] Collection of umbilical cord and transportation: the same method and requirements as in Example 1.

[0053] Washing and disinfection of the umbilical cord: with the method and requirements of Example 1, only 75% alcohol disinfectant is used.

[0054] Remove blood vessels and separate Wharton's jelly: Transfer the cleaned umbilical cord from the centrifuge tube to a petri dish, cut off 1 cm from the head and tail and discard, and cut the rest into 2 cm sections. Take a section of umbilical cord, peel off the outer skin with toothed forceps, remove the veins and arteries, and peel off the Wharton's jelly, taking care not to peel off the epidermal tissue of the umbilical cord. Wharton's jelly was placed in a 50mL centrifuge tube and washed twice with cleaning solution.

[0055] Transfer Wharton's jelly to a new centrifuge tube, add serum-free medium to make it just submerge Wharton's jelly, use...

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Abstract

The invention discloses a method for obtaining MSCs (mesenchymal stem cells) from umbilical cord. The method comprises the following steps: washing the umbilical cord and sterilizing the umbilical cord with entoiodine and / or 75% alcohol, removing blood vessels, and separating Wharton's jelly; performing planking culture; exchanging a medium for culture and performing harvesting on the 14th day orwhen the cell confluence reaches 45%-55%; performing digestion and filtration. With the method, primary MSCs can be obtained more quickly in shorter than 15 days, operation is simpler, more primary MSCs are obtained with the same quantity of tissue blocks, pollution can be effectively avoided, and antibiotics need not be added in subsequent culture to prevent cell pollution.

Description

technical field [0001] The invention relates to the field of cell preparation, in particular to a method for obtaining mesenchymal stem cells from umbilical cords. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells derived from the early mesoderm and ectoderm. MSCs have functions such as immune regulation, promotion of hematopoietic recovery, ability to repair damaged or diseased tissues and organs, and multi-lineage differentiation potential, so they have broad application prospects. [0003] However, the current method for preparing mesenchymal stem cells from the umbilical cord is time-consuming, the number of mesenchymal stem cells obtained is small, and it is easy to be polluted during the operation. [0004] For example, CN105586309A discloses a method for preparing mesenchymal stem cells from the umbilical cord. The umbilical cord after cleaning with antibiotic-added cleaning solution is cut into small sections with a len...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N2509/00
Inventor 林雨国周亚楠
Owner 厦门博瑞思生物科技有限公司
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