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Vitamin C induced mesenchymal stem cell membrane and preparation method thereof

A technology of mesenchymal stem cells and cell membranes, applied in the field of vitamin C-induced mesenchymal stem cell membranes and its preparation, can solve the problems of complex and time-consuming cell membranes, and achieve the effect of regeneration

Inactive Publication Date: 2012-01-11
王松灵 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the preparation of cell membranes is relatively time-consuming, complicated, and requires special temperature-sensitive materials such as PIPA-Am.

Method used

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  • Vitamin C induced mesenchymal stem cell membrane and preparation method thereof
  • Vitamin C induced mesenchymal stem cell membrane and preparation method thereof
  • Vitamin C induced mesenchymal stem cell membrane and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of periodontal ligament stem cell sheet

[0037] 1. Cell culture

[0038]Periodontal ligament stem cells can be obtained through purchase, and the purchase method is as described above. If it is obtained by culture, the culture method is as follows: immediately put the discarded tooth extracted by the patient into a sterile centrifuge tube filled with pre-cooled PBS, transfer it into the cell chamber, and isolate and culture periodontal ligament stem cells within 24 hours. Gently peel off the periodontal tissue around the teeth, take the periodontal tissue in the middle, wash it repeatedly with PBS, cut it into pieces, put it in the digestive solution containing type Ⅰ collagenase (3g / L) and Dispase (4g / L), at 37°C Digest for 1 hour, collect the cells through a 70 μm cell sieve, centrifuge at 1000 rpm for 10 minutes, and resuspend with culture medium to form a single cell suspension. Inoculate the cells in a 25cm2 cell culture flask in α-MEM medi...

Embodiment 2

[0109] Example 2 Preparation of Bone Marrow Mesenchymal Stem Cell Sheets

[0110] 1. Cell culture

[0111] Bone marrow mesenchymal stem cells can be purchased from cell banks, or can be obtained by culturing medical waste: bone marrow 5-10ml, diluted with an equal volume of 15% FBSα-MEM medium (containing 5mg heparin sodium). Using the whole bone marrow culture method, centrifuge the bone marrow α-MEM mixture at 1000rpm for 10min, discard the supernatant and fat mass, add α-MEM medium (containing 15% fetal bovine serum, 2mmol / L glutamine, 100U / ml penicillin and 100μg / ml streptomycin) in 20ml resuspended, pipet and mix well, inoculate in a culture bottle, place in a 37°C, 5% CO2 incubator for culture, perform a half-quantity liquid change on the third day, discard non-adherent cells, and then Change the medium every three or four days. Until the cells grew to 80% confluence, they were digested and passaged with 0.25% trypsin at a ratio of 1:2.

[0112] 2. Bone marrow mesench...

Embodiment 3

[0117] Embodiment 3 umbilical cord mesenchymal stem cell membrane culture method

[0118] 1. Cell culture

[0119] Take the discarded umbilical cord tissue of the newborn baby, soak it in α-MEM medium, take out the umbilical cord in the ultra-clean bench, fully wash the residual blood with D-Hank's balance solution, remove the umbilical vein, umbilical artery and umbilical cord adventitia, and peel off the Wharton glue. And shred it into 1mm×1mm×1mm tissue pieces. Move the tissue block into 0.5% collagenase II, add a little α-MEM, and digest it in a constant temperature oscillator at 37°C until the Wharton glue is completely digested. Aspirate the digestive solution containing cells into a sterile centrifuge tube, dilute it with 30 times the volume of α-MEM repeatedly, centrifuge at more than 2,000 rpm for 10 minutes, discard the supernatant, and culture with α-MEM containing 15% fetal bovine serum The base was washed to make the cell suspension evenly distributed, transferr...

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Abstract

The invention relates to a mesenchymal stem cell membrane, and concretely relates to a vitamin C induced mesenchymal stem cell membrane and a preparation method thereof. The preparation method which is simple, fast, and harmless has very important and far-reaching meanings on the tissue regeneration of the mesenchymal stem cell membrane in application fields of the cell substitution treatment and regeneration medical science, tissue engineering science and the like. The vitamin C induced mesenchymal stem cell membrane has a wide application prospect with continuous exploration and research and further advanced understanding of the mesenchymal stem cell membrane.

Description

technical field [0001] The invention relates to a mesenchymal stem cell membrane, in particular to a vitamin C-induced mesenchymal stem cell membrane and a preparation method thereof. Background technique [0002] In the traditional concept, the research field of tissue engineering technology mainly includes three aspects: seed cells, scaffold materials, and growth factors. Although the rapid development of biomaterial technology in recent years has made this classic construction strategy more and more mature, some disadvantages of this method have also emerged, and more and more scholars have begun to realize the importance of tissue construction by simulating embryonic development. [0003] In fact, the renewal and differentiation of stem cells are closely related to the microenvironment in which they live. The so-called microenvironment, simply speaking, is the dynamic environment in which cells live in the body, which is composed of cells, growth factors and extracellul...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 王松灵魏福兰施松涛范志朋丁刚
Owner 王松灵
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