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

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Problems solved by technology

But so far, the preparation of cell membranes is relatively time-consuming, com

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

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[0036] Example 1 Preparation of Periodontal Ligament Stem Cell Patch

[0037] 1. Cell culture

[0038] Periodontal ligament stem cells can be obtained through purchase, please refer to the above-mentioned way of purchase. If it is obtained by culture, the culture method is as follows: immediately put the discarded teeth of the patient into a sterile centrifuge tube containing pre-cooled PBS, transfer to the cell compartment, and separate and culture periodontal ligament stem cells within 24 hours. Gently peel off the periodontal tissue around the teeth, take the middle periodontal tissue, repeatedly wash with PBS, cut into pieces, and place it in a digestive solution containing type I collagenase (3g / L) and Dispase (4g / L) at 37°C After digestion for 1 hour, the cells were collected through a 70μm cell sieve, centrifuged at 1000 rpm for 10 min, and resuspended in the culture medium to form a single cell suspension. Inoculate the cells in a 25cm2 cell culture flask, in a-MEM medium...

Example Embodiment

[0109] Example 2 Preparation of Mesenchymal Stem Cell Membrane Sheet

[0110] 1. Cell culture

[0111] Bone marrow-derived mesenchymal stem cells can be purchased through cell banks, or cultured from 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 1000 rpm for 10 minutes, discard the supernatant and cellulite, and add α-MEM medium (containing 15% fetal bovine serum, 2mmol / L glutamine, 100U / ml penicillin and 100μg / ml streptomycin) was resuspended in 20ml, pipetted and mixed, then inoculated into a culture flask, placed in a 37°C, 5% CO2 incubator, cultured in a 5% CO2 incubator, and changed half of the medium on the third day, discarding non-adherent cells. Change the fluid every three or four days. Until the cells grow to 80% confluence, they are digested and passaged with 0.25% trypsin at 1:2.

[0112] 2. Preparation of Me...

Example Embodiment

[0117] Example 3 Umbilical cord mesenchymal stem cell patch culture method

[0118] 1. Cell culture

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

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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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