Application of human umbilical cord mesenchymal stem cell in preparation of drug for preventing Parkinson's disease or treating early Parkinson's disease
A technology of mesenchymal stem cells and Parkinson's disease, applied in the application field of medicine, can solve the problems of high cost, complicated differentiation steps, long cell preparation cycle, etc., and achieve the effect of easy acquisition
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Embodiment 1
[0052] Human umbilical cord mesenchymal stem cells were purchased from Saiye Biotechnology Company.
[0053] The purchased MSCs were planted in a 10cm-diameter petri dish, and 5mL of medium was added. Medium composition: containing DMEM / F12, 10% FBS, 0.1% penicillin and 0.1% streptomycin sulfate.
[0054] Subculture of human umbilical cord mesenchymal stem cells
[0055] (1) Add 1mL of 0.25% trypsin solution, quickly rotate the culture dish to completely cover the bottom of the dish, put it in a 37°C incubator and let it stand for 60-90s.
[0056] (2) Observing under a microscope, when most of the cells retract and become round, 1 mL of complete culture medium can be added to terminate the digestion.
[0057] (3) Gently pipette the cells several times until most of the cells are blown up to float, transfer the cell suspension to a 15mL centrifuge tube, centrifuge at 1200r / min for 5min.
[0058] (4) Discard the supernatant, add 1 mL of complete medium, gently pipette to susp...
Embodiment 2
[0061] Mouse tail vein injection of MSC suspension
[0062] Male BLC57 / 6 mice were used. The groups were divided into sham operation Sham group (not treated with 6-OHDA modeling but injected with saline in the substantia nigra), 6-OHDANS group (treated with 6-OHDA and injected with saline intravenously), 6-OHDAMSC group (6 -OHDA modeling treatment and vein transplantation of MSCs). Unilateral injection of normal saline mice and unilateral 6-OHDA Parkinson's disease model mice were purchased from Victoria Lihua Company.
[0063] On the second day after mice were established with 6-OHDA, MSCs were injected into the tail vein. The specific injection method is as follows:
[0064] Make a simple mouse holder: Take a 50ml centrifuge tube and drill a small hole at the bottom of the centrifuge tube for the mouse to breathe. Open a circular hole with a diameter of about 0.5 cm in the center of the lid of the centrifuge tube, so that the tail of the mouse can pass through. Use medi...
Embodiment 3
[0068] The Sham group, 6-OHDANS group, and 6-OHDAMSC group treated in Example 2 were injected with normal saline or transplanted with MSCs 3 days, 7 days, and 14 days after the behavioral function of the mice was evaluated. The specific mouse behavior tests were as follows:
[0069] (1) 3 days, 7 days, and 14 days after injection of normal saline or transplantation of MSCs, conduct behavioral tests.
[0070] (2) Weigh each mouse, inject 0.5 mg / kg apomorphine intraperitoneally, put it into a 2L beaker, and start timing for 5 minutes after each mouse is injected.
[0071] (3) Count the number of contralateral and ipsilateral turns of each mouse in 1 minute, and use a video camera to record the whole process. If the number of contralateral turns in 1 minute is ≥ 7, it is considered to have reached the standard of Parkinson's disease.
[0072] (4) Each mouse was counted 6-8 times, and the mouse was put back into the cage 30 minutes after the counting was completed.
[0073] The r...
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