Serum-free umbilical cord mesenchymal stem cell composition and application thereof

A mesenchymal stem cell and composition technology, applied in the field of umbilical cord mesenchymal stem cell composition, can solve the problems of joint function decline, poor treatment effect of articular cartilage injury, limited source of cartilage particles, etc., to achieve easy operation and avoid surgical risks Effect

Inactive Publication Date: 2019-08-09
苏州元复生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cartilage tissue generated after microfracture surgery is fibrocartilage, not hyaline cartilage. Therefore, although microfracture surgery has a certain effect in the short term, long-term follow-up shows that the joint function of microfracture surgery will still decline.
[0006] At the same time, the repair technique of filling the articular cartilage with its own cartilage particles can replace the patient's articular cartilage loss. 2 The treatment effect of articular cartilage injury is not good, and this treatment method has no therapeutic effect on subchondral bone, joint synovium and joint ligament

Method used

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  • Serum-free umbilical cord mesenchymal stem cell composition and application thereof
  • Serum-free umbilical cord mesenchymal stem cell composition and application thereof
  • Serum-free umbilical cord mesenchymal stem cell composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Isolation, purification and culture of umbilical cord mesenchymal stem cells

[0067] (1) Wash the umbilical cord tissue (remove blood cells): add an equal amount of normal saline to the centrifuge tube containing the umbilical cord, tighten the cap, shake for 3 minutes to fully wash the umbilical cord tissue, then stand still for 3-5 minutes to separate the different phases, and suck off the lower layer Water phase; repeat the above operation three times until the lower layer is relatively clear.

[0068] (2) Ultrasonic microwave pulverization: After absorbing and discarding the normal saline, add preheated DMEM equal to the volume of the umbilical cord, put it in a constant temperature microwave pulverizer, and conduct ultrasonic microwave pulverization at 37°C.

[0069] (3) Collect the precipitate: after digestion, centrifuge at 2000rpm for 10min, discard the digested umbilical cord in the upper layer, collect the bottom layer of the two tubes into a new centrifuge t...

Embodiment 2

[0074] Identification of umbilical cord mesenchymal stem cells

[0075] The P2 generation umbilical cord mesenchymal stem cells cultured in Example 1 were centrifuged and resuspended with 15% human albumin solution and 0.9% sodium chloride solution, wherein the volume of the human albumin solution and the sodium chloride solution The ratio was 0.1:1; the cell concentration was adjusted to l×10 after cell counting 8 / L, respectively reacted with human anti-CD73, CD90, CD34, CD45, MHC-II monoclonal antibodies at room temperature for 30 minutes, resuspended the cells in PBS, and detected them by flow cytometry. The test results are shown in the table below and figure 1 . The cell surface antigen marker expression of umbilical cord mesenchymal stem cells was analyzed by flow cytometry, and the cells were of high purity.

[0076] surface antigen CD73 CD90 CD34 CD45 MHC-II result 99.06％ 98.34％ 0.57％ 3.43％ 1.87％

Embodiment 3

[0078] Umbilical cord mesenchymal stem cells osteogenic, adipogenic, chondrogenic ability test

[0079] The cells cultured in Example 1 were used as the group of the present invention, and the osteogenic, adipogenic and chondrogenic abilities were tested. Alizarin Red S, Oil Red O, and Alcian blue staining showed that the cells cultured in Example 1 had the ability to differentiate into bone, fat, and cartilage in vitro after 3-4 weeks of culture. see results figure 2 , reflecting the in vitro differentiation ability of umbilical cord mesenchymal stem cells

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Abstract

The invention discloses a serum-free umbilical cord mesenchymal stem cell composition and application thereof. The composition comprises umbilical cord mesenchymal stem cells, a human albumin solutionand a sodium chloride solution, wherein the mass fraction of the sodium chloride solution is 0.9%, and the umbilical cord mesenchymal stem cell composition is applied to preparing medicines for treating or preventing osteoarthritis. In this way, according to the umbilical cord mesenchymal stem cell composition and application thereof, an optimized preparation volume and a component concentrationare adopted, the umbilical cord mesenchymal stem cells are combined with sodium chloride to prepare an injection, and osteoarthritis injury can be treated under the condition that the low injection amount is about 3-6 ml.

Description

technical field [0001] The invention relates to the technical field of osteoarthritis treatment, in particular to a serum-free umbilical cord mesenchymal stem cell composition and its application. Background technique [0002] The incidence of Osteoarthritis (OA) ranks second among chronic diseases, second only to cardiovascular diseases. At present, there are 360 ​​million patients in the world, and the affected population in mainland China is about 120 million. The prevalence rate can reach 50% in the crowd over 60 years old, and 80% in the crowd over 75 years old. [0003] Hyaline cartilage and synovial fluid together cushion and lubricate the joints. Articular hyaline cartilage has poor repair ability and cannot spontaneously heal or regenerate when it is damaged by external shocks. Hyaline cartilage does not contain blood, nerves, and lymphatic system, so it cannot stimulate the body's repair response. [0004] The symptoms of early osteoarthritis are not obvious eno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K9/08A61K47/42A61P19/02A61P19/08A61P29/00C12N5/0775
CPCA61K35/28A61K9/08A61K9/0019A61K47/42A61P19/02A61P19/08A61P29/00C12N5/0665C12N2509/00C12N2501/998
Inventor 汪文
Owner 苏州元复生物科技有限公司
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