A double-sided affinity-modified scaffold-stem cell composite material
A technology of combining materials and nucleus pulposus cells, which is applied in the field of biological tissue engineering cell carrier materials, can solve the problems of stem cell modification, reduce the induction effect of biological materials, and achieve the effect of avoiding damage
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Embodiment 1
[0026] Example 1: It is intended to use 500 μL of 6 mg / mL type II collagen stock solution to construct a double-sided affinity-modified pseudo-nucleus pulposus scaffold-stem cell composite material.
[0027] see figure 1 , the construction method is as follows: (1) 500 μL of 6 mg / mL type II collagen storage solution and 162 μL of 50 mg / mL chondroitin sulfate storage solution were mixed on ice, then 100 μL of 10×PBS was added, and the pH was adjusted to 7.2 with 1M NaOH, Dilute to volume with high glucose medium to obtain 1mL type II collagen / chondroitin sulfate mixture. The final concentration of each substance is: 3mg / mL type II collagen, 8.1mg / mL chondroitin sulfate, 1×PBS. The mixture was allowed to stand at 37°C for 1 hour to form a gel, and the gelled type II collagen / chondroitin sulfate gel was freeze-dried overnight at -80°C to obtain a traditional type II collagen / chondroitin sulfate bioscaffold material. (2) 4×10^5 nucleus pulposus cells were resuspended in 1 mL of ...
Embodiment 2
[0028] Example 2: It is intended to use 4×10^5 adipose-derived mesenchymal stem cells to construct a bilateral affinity-modified pseudo-nucleus pulposus scaffold-stem cell composite material.
[0029] see figure 1 , and the construction method is as follows: (1) 1 mL of 6 mg / mL type II collagen stock solution and 324 μL of 50 mg / mL chondroitin sulfate stock solution were mixed on ice, and then 200 μL of 10×PBS was added, and the pH was adjusted to 7.2 with 1M NaOH, and then used The cell high glucose medium was adjusted to volume to obtain 2mL type II collagen / chondroitin sulfate mixture. The final concentration of each substance is: 3 mg / mL type II collagen, 8.1 mg / mL chondroitin sulfate, 1×PBS. The mixture was allowed to stand at 37°C for 1 hour to form a gel, and the gelled type II collagen / chondroitin sulfate gel was freeze-dried overnight at -80°C to obtain a traditional type II collagen / chondroitin sulfate bioscaffold material. (2) Take 8×10^5 nucleus pulposus cells an...
Embodiment 3
[0030] Example 3: In vitro related evaluation of the double-sided affinity-modified nucleus pulposus scaffold-stem cell composite material.
[0031] After obtaining the nucleus pulposus scaffold modified by the nucleus pulposus cells through the steps described above, we detected the surface structure of the pseudonucleus pulposus scaffold by scanning electron microscopy, and measured the elastic modulus of the pseudonucleus pulposus scaffold by rheological testing. Proved that it has good microstructure and physical properties ( figure 2 ). After ManNProp was synthesized by mannosamine and propionyl chloride, we verified the synthetic results of ManNProp by NMR analysis ( image 3 ). And the cytotoxicity of ManNProp was measured, it was found that ManNProp has no toxic effect on stem cells at a concentration of 0.1 μmol / ml ( Figure 4 ). After targeted editing of stem cells with ManNProp, we performed mass spectrometry analysis on the glycosyl groups on the surface of st...
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