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

Active Publication Date: 2019-09-06
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, there are few studies in the midbrain and Parkinson's disease, and they mainly focus on the research on the damage and apoptosis of dopaminergic neurons caused by neuroinflammation and the protective effect of neuroprotective factors on astrocytes. Less research has been done in iron metabolism
In vitro studies, there are no reports on the interaction of primary cultured astrocytes and ventral mesencephalon (VM) neurons co-culture system for iron transport and iron metabolism

Method used

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  • Applications of primary astrocyte/primary midbrain ventral neuron co-culture system in iron metabolism
  • Applications of primary astrocyte/primary midbrain ventral neuron co-culture system in iron metabolism
  • Applications of primary astrocyte/primary midbrain ventral neuron co-culture system in iron metabolism

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention provides applications of a primary astrocyte/primary midbrain ventral neuron co-culture system in iron metabolism, especially in iron transport, and belongs to the field of cytobiology.The technical schemes detect a co-culture system by designing test methods, the sources of iron in neurons, the mutual influence between interstellar astrocyte and the directionality problems of irontransport can be cleared at the cellular level. The established primary astrocyte and primary midbrain ventral neuron co-culture system is a novel cell model for studying Parkinson diseases, and can be used for studying the related research of high iron in the brain of neurodegenerative diseases such as the Parkinson diseases.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to the application of a primary astrocyte / primary midbrain ventral neuron co-culture system in iron metabolism, especially iron transport. Background technique [0002] Parkinson's disease (PD) is the second most common neurodegenerative disease in middle-aged and elderly people, and its pathological feature is the loss of dopamine (DA) neurons in the substantia nigra. In recent years, the impairment of the function of DA neurons in the substantia nigra-striatum system caused by high iron in the brain has become a hot issue that has attracted more and more attention from neuroscientists. At present, the research on iron deposition in PD mainly focuses on DA neurons. When iron is loaded, neurons, microglia and oligodendrocytes have iron deposition, but astrocytes do not have iron deposition under the same conditions. . Then, how astrocytes play a role in brain iron metabolis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079A61K35/30A61P25/16
CPCA61K35/30A61P25/16C12N5/0622C12N2502/081C12N2509/00
Inventor 谢俊霞王俊许曼曼
Owner QINGDAO UNIV
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