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Efficient preparation method of placental mesenchymal stem cells through homogenizer

A technology of stem cells and homogenizer, applied in the field of mesenchymal stem cells and their separation and purification, can solve the problems of low harvest, immune rejection, clinical application limitations, etc., and achieve broad clinical application prospects, simple expansion and culture, low cost The effect of immunogenicity

Inactive Publication Date: 2019-10-25
陈蓓宁 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most studied is mesenchymal stem cells derived from bone marrow. Mesenchymal stem cells have also been found in other tissues including fat, muscle, nerve, and dental pulp, but their acquisition and isolation are more difficult than those of bone marrow.
However, obtaining mesenchymal stem cells from these tissues will cause certain trauma to the donor, and most donors will have concerns
Moreover, the mesenchymal stem cells obtained from these highly differentiated tissues usually have a certain degree of antigenicity, and there will be immune rejection in allogeneic transplantation, and the amount of harvest is small, which requires a lot of time and money for in vitro expansion and culture , so it will be greatly limited in clinical application

Method used

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  • Efficient preparation method of placental mesenchymal stem cells through homogenizer
  • Efficient preparation method of placental mesenchymal stem cells through homogenizer
  • Efficient preparation method of placental mesenchymal stem cells through homogenizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Transportation of placenta

[0025] The placenta taken out during normal delivery or caesarean section is immediately put into the placenta storage solution. The placenta storage solution is DMEM medium containing 10% fetal bovine serum and four antibiotics. Fetal bovine serum is Australian bovine fetal bovine serum (Gibco). Antibiotics are 50-150IU / mL penicillin, 60-130μg / mL streptomycin, 20-50μg / mL gentamicin and 1-2.5μg / mL amphotericin B. Brought back to the laboratory by the hospital in a 4°C environment for subsequent treatment.

Embodiment 2

[0026] Example 2 Cleaning of the placenta

[0027] Remove the placenta from the transport solution, place it in a large sterile beaker, and add a PBS balanced salt solution containing four antibiotics. The antibiotics are 50-150IU / mL penicillin, 60-130μg / mL streptomycin, 20-50μg / mL gentamicin and 1-2.5μg / mL amphotericin B. Under aseptic conditions, gently squeeze and wash repeatedly by hand to thoroughly wash away the residual blood in the placenta. The placenta washes to pale pink.

Embodiment 3

[0028] Example 3 Clipping of placental lobules

[0029] Place the cleaned placenta in a sterile tray, and cut the placenta lobules with sterilized surgical scissors. After washing again with PBS balanced salt solution containing antibiotics, placental lobules were placed in DMEM medium containing 5-10% fetal calf serum and four antibiotics for 5 minutes to maintain their activity.

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Abstract

The invention relates to separation and purification of placental derived mesenchymal stem cells and belongs to the field of stem cell and tissue engineering. The efficient preparation method includes: with the consent of the parturient and her family, placing the placenta after caesarean birth of the parturient in a storage solution with a variety of antibioctics for a short period of time; crushing the placental after repeatedly cleaning blood and water of the placental in a laboratory, digesting by collagenase and patting with a homogenizer to obtain a cell suspension; using a density gradient centrifugation method to obtain the placental mesenchymal stem cells. The method for separating the placental derived mesenchymal stem cells with the homogenizer is multiple times higher than thatof traditional methods, the hydrolysis time is reduced, and the success rate is improved; the obtained mesenchymal stem cells have similar amplification capability and multifunctional differentiationpotential with bone marrow derived mesenchymal stem cells in vitro and are potential seed cell sources for tissue engineering and cell therapy, thereby having important clinical significance.

Description

[0001] Invention field [0002] The invention relates to placenta-derived mesenchymal stem cells and separation and purification thereof, and belongs to the field of stem cells and tissue engineering. Background technique [0003] In recent years, the primary problem facing the continued popularity of tissue engineering and cell therapy is the source of seed cells. Because stem cells have a certain self-renewal ability in vitro, and they have the potential to differentiate into one or more types of cells under specific induction conditions, and even all types of cells in the human body, they have gradually attracted the attention of scientific researchers, and they happen to be able to solve the problem. The source of tissue cells is scarce and expansion is difficult, and it is easy to lose phenotype during in vitro culture. Therefore, stem cells show good clinical application prospects. According to the current research reports, there are two types of stem cells successfully sep...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2509/00
Inventor 陈蓓宁邢为藩张鹏
Owner 陈蓓宁
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