A kind of anti-melanoma stem cell patch and preparation method thereof
An anti-melanoma and stem cell technology, applied in the field of biotechnology and regenerative medicine, can solve the problems of difficult to achieve radical cure and little effect of malignant melanoma treatment, and achieve the effects of promoting apoptosis, increasing interferon synthesis, and enhancing killing effect.
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specific Embodiment 1
[0025] Specific embodiment 1: as figure 1 Shown is a flow chart of the preparation steps of an anti-melanoma stem cell patch of the present invention, and the described method comprises the following steps:
[0026] Step 1: Preparation of bone marrow-derived mesenchymal stem cells
[0027] The femoral bone marrow of spontaneously aborted fetuses was collected under sterile conditions (human embryonic bone marrow-derived mesenchymal stem cells used in the experiment were all spontaneously aborted fetuses, and the mothers of the aborted fetuses were informed and signed the consent form agreeing that the aborted fetuses were used for stem cell research), and used 20% fetal bovine Wash the bone marrow of the femur with the DMEM medium (containing penicillin 100 / mL, streptomycin 100 μg / mL, pH7.4) and pour it into a plate, and use a syringe to suck and blow properly to make the bone marrow tissue cells basically evenly dispersed, and then count , to 1 x 10 4 / cm 2 Density inocu...
specific Embodiment 2
[0041] Specific Example 2: Flow cytometric identification of mesenchymal stem cells derived from bone marrow
[0042] The isolated human embryonic bone marrow-derived mesenchymal stem cells were detected by flow cytometry: more than 95% of CD166-positive cells and less than 3% of CD34-positive cells (CD166 is a characteristic surface marker of BMSC cell membrane, and BMSC cell membrane does not express CD34 ), the cell viability detected by trypan blue was above 95%.
Embodiment 3
[0043] Embodiment 3 animal test
[0044] 1. Preparation of melanoma mouse model
[0045] ①The preparation concentration is 1×10 7 cells / ml B16 melanoma cell suspension; ②Use a dermatome to remove the back hair of 6-8 week-old C57BL / 6J mice, and the hair removal area is about 4cm 2 ; ③ B16 melanoma cells were subcutaneously injected into the back of 40 mice, 0.2ml each; the tumor growth was observed the next day, the size of the induration was measured, the survival period of the mice was observed, and relevant data were recorded.
[0046] 2. Treatment of Melanoma Mice
[0047] One week later, the tumor-bearing mice were randomly divided into two groups, 20 in each group. The experimental group had the successfully prepared stem cell patch applied to the tumor body, and the control group had no patch applied as a blank control. The patch was replaced every four days, and the treatment continued. 4 times. When replacing the patch, measure the length and width of the tumor w...
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