Composition having efficacy of promoting mesenchymal stem cells to secrete cytokine
A technology of mesenchymal stem cells and cytokines, applied in the field of compositions with the effect of promoting mesenchymal stem cells to secrete cytokines, can solve the problems of undisclosed induction compositions, etc., and achieve the effect of enhanced quantity
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Embodiment 1
[0028] This embodiment provides a composition for promoting the secretion of cytokines by mesenchymal stem cells and its application. Specifically, the composition is added to high-sugar DMEM to prepare adipose-derived mesenchymal stem cell secretion cytokine induction medium, which is used to stimulate adipose-derived mesenchymal stem cells to secrete cytokines.
[0029] 1. Adipose-derived mesenchymal stem cell secretion cytokine induction medium (see Table 1): 500mL high-glucose DMEM, 0.65g VC, 3g insulin, 0.5g trisodium citrate, 0.55g hyaluronic acid.
[0030] 2. Preparation of Adipose-derived Mesenchymal Stem Cell Factors:
[0031] (1) After the adipose-derived mesenchymal stem cells grow to about 85%, they are subcultured in LONZA serum-free fat medium;
[0032] (2) Transfer P3-P5 cells to a cell culture dish filled with LONZA serum-free fat medium, culture until the cell growth density reaches 50%-65%, discard the original medium, and wash three times with PBS , replac...
Embodiment 2
[0037] This embodiment is a modification example of Embodiment 1, and the changes relative to Embodiment 1 mainly include:
[0038] 1. Adipose-derived mesenchymal stem cells secrete cytokine-inducing medium formula. The medium formula in this example (see Table 1) is as follows: 500mL high-sugar DMEM, 0.15gVC, 0.53g insulin, 0.1g trisodium citrate , 0.25g hyaluronic acid.
[0039] 2. In step (2), transfer the P3~P5 cells to a cell culture dish filled with LONZA serum-free fat medium, cultivate until the cell growth density reaches 50%~65%, discard the original medium, Wash three times with PBS, replace with adipose-derived stem cell secretion cytokine induction medium for culture, after 1h, the cells were subjected to 37°C, 5% CO 2 , 10%O 2 Hypoxic treatment, observe the growth status of the cells every 12, and culture continuously for 84 hours;
[0040] Other operations are the same as in Example 1.
Embodiment 3
[0042] This embodiment is a modification example of Embodiment 1, and the changes relative to Embodiment 1 mainly include:
[0043] 1. Adipose-derived mesenchymal stem cells secrete cytokine-inducing medium formula. The medium formula in this example (see Table 1) is as follows: 500mL high-glucose DMEM, 1gVC, 5g insulin, 1g trisodium citrate, 1g transparent uric acid.
[0044] 2. In step (2), transfer the P3~P5 cells to a cell culture dish filled with LONZA serum-free fat medium, cultivate until the cell growth density reaches 50%~65%, discard the original medium, Wash three times with PBS, replace with adipose stem cell secretion cytokine induction medium for culture, after 3h, the cells were subjected to 37°C, 5% CO 2 , 3%O 2 Hypoxic treatment, observe the growth status of the cells every 12, and continuously culture for 60 hours;
[0045] Other operations are the same as in Example 1.
[0046] Table 1
[0047]
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