Application of amniotic epithelial cells and cell preparations thereof
A technology for amniotic epithelial cells and cell preparations, applied in the field of biomedicine, can solve problems such as the problem of neurogenic bladder treatment, and achieve the effect of restoring physiological functions and having a wide range of sources.
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Embodiment 1
[0052] Example 1. hAEC isolation and culture
[0053] Take the placenta of healthy caesarean section pregnant women, and strip the amniotic membrane from the chorion. Rinse with PBS to remove blood cells, digest with 0.25% trypsin at 37°C for 30 min, pipette, add RPMI1640 containing 10% FBS to stop the digestion, filter through a 100-mesh sieve, centrifuge at 1,500 rpm, discard the supernatant, and dilute the mixture with 1×10 4 / cm 2 Inoculate in 10cm dish. hAECs were placed in RPMI-1640 (10% FBS, 100 μg / ml streptomycin, 100 U / ml penicillin, 10 ng / ml EGF and 0.3 mg / ml glutamine) at 37°C, 5% CO 2 cultured in a cell culture incubator.
Embodiment 2
[0054] Example 2. Preparation of hAEC cell preparation
[0055] Example 1 hAEC cultured for 24-72 hours, remove the medium, digest the adherent hAEC with 0.25% trypsin and centrifuge, then rinse with 2ml of PBS solution and centrifuge again, resuspend in PBS and adjust the cell concentration to 105 / μl for later use, namely hAECs cell preparations were obtained.
Embodiment 3
[0056] Example 3.hAECs Treat Neurogenic Bladder Animals
[0057] 1. Animals
[0058] SPF grade female adult Sprague-Dawley rats were used in the experiment, purchased from Shanghai Slack Experimental Animal Center, weighing 250-300g, reared in a single cage, free to ingest water and food, and lighted to form a day / night cycle of 12-12h. Experimental animals were divided into control group, operation (spinal cord injury) group and treatment (cell transplantation) group.
[0059] 2. Surgical method
[0060] The animals in the operation group were anesthetized by intraperitoneal injection of sterile 10% chloral hydrate (400mg / kg), lying prone, making an incision about 2 cm long centered on the thoracic 12 spine, cutting the skin, and bluntly peeling off the fascia layer by layer, Muscles, after the spinous process and lamina are exposed, the thoracic 11 spinous process and lamina are bitten off, and the spinal cord and central vein are exposed. The exposed spinal cord was full...
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