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Placenta mesenchymal stem cell injection, preparation method and application thereof

A kind of stem cell and injection technology, applied in the field of stem cells, can solve the problems of low immunogenicity and less application, and achieve the effects of low immunogenicity, strong proliferation ability, and convenient material acquisition.

Inactive Publication Date: 2017-07-07
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the continuous development of stem cell biology research, MSC has biological characteristics such as low immunogenicity and strong immune regulation (can evade immune recognition, suppress immune response), inflammatory chemotaxis, tissue repair, etc. Injection of MSC can improve the immune tolerance after transplantation and prolong the survival time of the graft. It has also been effectively used in the treatment of graft-versus-host disease, especially in the treatment of immune rejection such as kidney transplantation and bone marrow transplantation. effect, but it is less used in the treatment of acute rejection after liver transplantation

Method used

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  • Placenta mesenchymal stem cell injection, preparation method and application thereof
  • Placenta mesenchymal stem cell injection, preparation method and application thereof
  • Placenta mesenchymal stem cell injection, preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0072] Embodiment 1 Preparation method of placental mesenchymal stem cells

[0073] 1. Primary isolation of placental mesenchymal stem cells

[0074] Under sterile conditions, take the fresh placenta discarded after delivery, peel off the placenta, wash it with PBS buffer solution, and cut the placenta tissue into 5×5cm 2 , add 3-5 times the volume of 0.25% trypsin to digest, place on a 200r / min 37°C constant temperature shaker, digest for 30min, and shake vigorously several times every 10min to remove epithelial cells.

[0075] After the digestion is complete, transfer it to a storage bottle, add 150 mL of PBS buffer solution, shake vigorously, and repeat this operation 2 times. Transfer placental tissue to a small beaker and chop to 1mm 3 . Then transfer to a storage bottle, add 20 mL of 0.5% type I collagenase and complete medium (high glucose DMEM+10% FBS), so that the final concentration of collagenase is 0.1-0.2%. Place in a constant temperature shaker at 37°C, 250r / ...

Embodiment 2

[0088] The preparation of embodiment 2 placental mesenchymal stem cell injection

[0089] Prepare 100mL stem cell injection: albumin concentration is 20% human serum albumin stock solution 15mL (mass volume ratio final concentration is 3%), vitamin C 0.5g (mass volume ratio final concentration is 0.5%), pearl ginsenoside 6g ( Mass volume ratio final concentration is 6%) and 0.9% sodium chloride injection 75mL are mixed, placental mesenchymal stem cells (cell suspension after recovery is 10mL, contains DMSO 1mL, FBS 9mL, cell final concentration 1×10 6 pcs / mL) in the mixture and resuspended to complete the preparation.

[0090] The reserved samples of the placental mesenchymal stem cell injection prepared above are tested for endotoxin, bacteria and fungi, and mycoplasma. If the test results are all negative, it is qualified.

Embodiment 3

[0091] The preparation of embodiment 3 placental mesenchymal stem cell injection

[0092] Prepare 100mL stem cell injection: albumin concentration is 20% human serum albumin stock solution 5mL (mass volume ratio final concentration is 1%), vitamin C 0.3g (mass volume ratio final concentration is 0.3%), pearl ginsenoside 2g ( Mass volume ratio final concentration is 2%) mixed with 0.9% sodium chloride injection 85mL, placental mesenchymal stem cells (cell suspension after recovery is 10mL, containing DMSO 0.5mL, FBS 4.5mL, cell final concentration 1× 10 6 pcs / mL) in the mixture and resuspended to complete the preparation.

[0093] The reserved samples of the placental mesenchymal stem cell injection prepared above are tested for endotoxin, bacteria and fungi, and mycoplasma. If the test results are all negative, it is qualified.

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Abstract

The invention relates to the technical field of stem cells and especially relates to a placenta mesenchymal stem cell injection, a preparation method and application thereof. The placenta mesenchymal stem cell injection comprises placenta mesenchymal stem cells, vitamin C, dimethyl sulfoxide (DMSO), fetal calf serum (FBS), human albumin, rhizoma panacis majoris saponin and solvent. The placenta mesenchymal stem cell injection prepared according to the invention is characterized in that the materials are conveniently taken and the ethical violation problem is avoided; the injection prepared according to the invention can effectively preserve the activity of stem cells, can be directly used for recovery, need not be cleaned and centrifuged and is conveniently used; after the injection prepared according to the invention is fed back, the differentiation of the mesenchymal stem cells to the hepatic cells and the maturation of the hepatic cells can be boosted and the GVHD occurrence rate can be reduced.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a placental mesenchymal stem cell injection and a preparation method and application thereof. Background technique [0002] The number of people infected with hepatitis B virus (HBV) in my country has reached 120 million, and the number of patients with chronic hepatitis is as high as more than 30 million. Among them, viral hepatitis B (hepatitis B) is the main cause of liver cirrhosis and primary hepatocellular carcinoma (liver cancer). The number of deaths due to liver cirrhosis, liver cancer, and liver failure is as high as 300,000 to 500,000, which seriously threatens people's health. For end-stage hepatitis, cirrhosis, and hereditary and metabolic liver diseases, orthotopic liver transplantation is currently the only sure and effective treatment, but there are various complications after liver transplantation, mainly including infection complications and rejection , bilia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K47/22A61K47/42A61K47/28A61K47/20A61K47/46A61K9/08A61P1/16A61P31/20
CPCA61K35/28A61K9/0019A61K9/08A61K47/20A61K47/22A61K47/28A61K47/42A61K47/46
Inventor 葛啸虎陈海佳王一飞李丽娟王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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