Cell Technology Topic: Human Umbilical Artery Smooth Muscle Cell Culture Experiment
Human umbilical arterial smooth muscle cell culture can: (1) obtain human umbilical arterial smooth muscle cells; (2) as a research substrate for major arterial diseases; (3) study vascular models; (4) continue to provide pharmacology and treatment for vascular diseases information.
experimental method
- Digestive method
- Patch method
| Principle of experimental method | Human umbilical arterial smooth muscle cells were cultured by digestive method, and morphological observation was performed with an inverted microscope and cultured cells were identified by immunohistochemistry. |
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| Experimental Materials | Umbilical cord |
| Reagents, kits | Collagenase Digestion Trypsin Collagenase Elastase DMEM |
| Instruments, consumables | Ophthalmic scissors dropper centrifuge Petri dish CO2 incubator |
| Experimental procedure | First, the experimental steps 1. Rinse in ice-cold sterile D-Hanks' solution, carefully remove the fat, capsule, blood vessels and other tissues with ophthalmic scissors, transfer to penicillin vials, add a small amount of sterile D-Hanks' solution, and cut into 0.1- with ophthalmic scissors. Fragments of 1.0 mm 3 size. 2. Use a dropper to gently aspirate the upper layer of small fat pieces and oil droplets, and then wash it 8 to 10 times with sterile D-Hanks' solution. 3. Add 10 volumes of sterile collagenase digestive solution (1 mg/mL), digest at room temperature for 30 min, centrifuge at 1000 rpm for 5 min, discard the supernatant, add 1 mg/mL trypsin for digestion at 37 °C for 20 min, centrifuge again. Min, discard the supernatant. 4. Add trypsin at 37 ° C for 20 min, centrifuge again for 5 min, discard the supernatant. 5. Add collagenase (1 mg/mL) elastase (0.5 mg/mL) and digest the digestion solution for 30 min at 37 °C, centrifuge for another 5 min, and discard the digestive juice. 6. Add the mixed digestive solution at 37 ° C for 20 min, centrifuge again for 5 min, resuspend the cells in DMEM medium containing 10% FBS, and transfer to a Petri dish at 37 ° C, 5% CO 2 incubator for cultivation. |
Waist massager
Waist massager
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