Application of primary astrocytes/primary ventral midbrain neurons co-culture system in the study of iron metabolism between astrocytes and neurons

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: 2021-05-28
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 is no report 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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  • Application of primary astrocytes/primary ventral midbrain neurons co-culture system in the study of iron metabolism between astrocytes and neurons
  • Application of primary astrocytes/primary ventral midbrain neurons co-culture system in the study of iron metabolism between astrocytes and neurons
  • Application of primary astrocytes/primary ventral midbrain neurons co-culture system in the study of iron metabolism between astrocytes and neurons

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 produc...

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 Biotechnology Research Institute.

[0057] 2.3 Experimental Instruments

[0058] Ultra-clean bench: Suzhou Purification Equipment Co., Ltd.; CO 2 Incubator: American Thermo Fisher Scientific product; ordinary ...

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.; CO2 Incubator: 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 VARIOKLAV...

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Abstract

The invention proposes the application of a primary astrocyte / primary midbrain ventral neuron co-culture system in the study of iron metabolism between astrocytes and neurons, and belongs to the field of cell biology. By designing an experimental method to test the co-culture system, the source of neuronal iron, its interaction with astrocytes, and the direction of iron transport can be identified at the cellular level. The co-culture system of primary astrocytes and primary midbrain ventral neurons established in the present invention is a new cell model for studying Parkinson's disease, and can be used to study the high iron in the brain of Parkinson's disease and other neurodegenerative diseases in related research.

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 the study of iron metabolism between astrocytes and neurons. 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 bra...

Claims

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Application Information

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Patent Type & Authority Patents(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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