Applications of primary astrocyte/primary midbrain ventral neuron co-culture system in iron metabolism
An astrocyte and co-culture system technology, applied in the field of cell biology, can solve the problem of less research on iron metabolism, and achieve the effects of reducing experimental consumption, improving research efficiency, and reducing costs
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Embodiment 1
[0035] Example 1 Primary culture of astrocytes
[0036] 1.1 Experimental animals
[0037] Newborn 24h Wistar suckling mice (provided by the Animal Center of Qingdao Institute for Drug Control) were selected as experimental animals for the primary culture of astrocytes.
[0038] 1.2 Experimental reagents
[0039]DMEM / F-12(1:1): product of Gibco; fetal bovine serum (FBS Gold): product of Gibco, EU grade; poly-lysine (poly-D-lysine-hydrobromide, poly-D-lysine) , boric acid, sodium tetraborate: products of Sigma Company; penicillin-streptomycin solution (100X), trypsin (Trypsin): products of Jiangsu Haimen Biyuntian Institute of Biotechnology.
[0040] 1.3 Experimental Instruments
[0041] Ultra-clean bench: Suzhou Purification Equipment Co., Ltd.; CO2 incubator: American Thermo Fisher Scientific product; ordinary centrifuge: Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.; incubator and oven: German MEMMERF product; -20°C and 4°C Refrigerator: Qingdao Haier produ...
Embodiment 2
[0052] Example 2 Primary Culture of Ventral Mesencephalon (VM) Neurons
[0053] 2.1 Experimental animals
[0054] As experimental animals, 14-day pregnant Wistar mice (provided by the Animal Center of Qingdao Institute of Drug Control) were selected, placed at a room temperature of 20° C., and lived under the conditions of 12:12 hours of day and night cycle light, free access to food and water.
[0055] 2.2 Experimental reagents
[0056] DMEM / F-12 (1:1): product of Gibco; B27: product of Gibco; fetal bovine serum (FBS Gold): product of Gibco, EU grade; poly-lysine (poly-D-lysine-hydrobromide, poly-D-lysine), boric acid, sodium tetraborate: products of Sigma Company; penicillin-streptomycin solution (100X), trypsin (Trypsin): products of Jiangsu Haimen Biyuntian Institute of Biotechnology.
[0057] 2.3 Experimental Instruments
[0058] Ultra-clean bench: Suzhou Purification Equipment Co., Ltd.; CO 2 Incubator: American Thermo Fisher Scientific product; ordinary centri...
Embodiment 3
[0067] Example 3 Establishment of co-culture system of astrocytes and VM neurons and intervention grouping
[0068] 3.1 Experimental reagents
[0069] DMEM / F-12 (1:1): product of Gibco; B27: product of Gibco; fetal bovine serum (FBS Gold): product of Gibco, EU grade; penicillin-streptomycin solution (100X), trypsin (Trypsin ): product of Jiangsu Haimen Beyantian Biotechnology Research Institute; 6-hydroxydopamine, 6-OHDA: product of sigma company; ascorbic acid: product of sigma company; FAC: product of sigma company.
[0070] 3.2 Experimental Instruments
[0071] Ultra-clean bench: Suzhou Purification Equipment Co., Ltd.; CO 2Incubator: American Thermo Fisher Scientific product; ordinary centrifuge: Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.; incubator and oven: German MEMMERF product; -20℃ and 4℃ refrigerator: Qingdao Haier product; -80℃ ultra-low temperature refrigerator: American Thermo Fisher Scientific product; high pressure sterilizer: German VARIOKLA...
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