Bionic three-dimensional nerve blood vessel unit direct contact co-culture system and construction method thereof

Abstract

The invention discloses a bionic three-dimensional nerve blood vessel unit direct contact co-culture system which at least comprises neurons, semipermeable membranes, three-dimensional cultured astrocytes, semipermeable membranes and vascular endothelial cells, the three-dimensional cultured astrocytes are attached to the non-vascular endothelial cell side of a semipermeable membrane-vascular endothelial cell complex, and the semipermeable membrane-vascular endothelial cell complex is attached to the non-vascular endothelial cell side of the semipermeable membrane-vascular endothelial cell complex. And the semi-permeable membrane is attached to the non-neuron cell side of the semi-permeable membrane-neuron complex to form a direct contact co-culture system of vascular endothelial cells, three-dimensional culture astrocytes and neurons. The invention further discloses a construction method of the bionic three-dimensional nerve blood vessel unit direct contact co-culture system. According to the bionic three-dimensional nerve and blood vessel unit direct contact co-culture system and the construction and culture method thereof provided by the invention, physiological functions of blood brain barrier, nutrition coupling, electrophysiological coupling and the like of the nerve and blood vessel unit in vivo can be comprehensively and effectively simulated.

Description

technical field [0001] The invention relates to the technical field of three-dimensional cell culture, in particular to a bionic three-dimensional neurovascular unit direct contact co-culture system and a construction method thereof. Background technique [0002] The neurovascular unit (NVU) is mainly composed of vascular endothelial cells (VEC), astrocytes and neurons (Neuron) figure 1 ). It mainly has physiological functions such as blood-brain barrier, nutritional coupling and electrophysiological coupling. [0003] Among them, vascular endothelial cells and astrocytes jointly participate in the formation of the blood-brain barrier (BBB), which can effectively limit the substances in the blood from entering the central nervous system. state plays an important role. [0004] The neurovascular unit has a trophic coupling function. In vivo, vascular endothelial cells, astrocytes and neurons form a multicellular complex structure in direct contact ( figure 1 ). Vascular...

AI Technical Summary

Problems solved by technology

[0007] First of all, the existing two-dimensional multicellular neurovascular unit direct contact co-culture system construction method is to mix various cells such as vascular endothelial cells, astrocytes and neurons together for co-culture, although it can form individual cells However, it is impossible to simulate the independent layered structure of the neurovascular unit in the body, so that the biological function of the neurovascular unit cannot be effectively simulated, and it is difficult to selectively detect the function and metabolism of a certain type of cell in the co-culture system Happening

Second, the existing direct contact co-culture system of transwell monolayer membrane can only prepare an independent layered structure system in which two layers of cells communicate directly, such as vascular endothelial cells-astrocytes, astrocytes- Neurons and other independent layered structures, the remaining cells need to be inoculated on the bottom of the culture dish to form indirect contact, and intercellular communication can only be carried out through indirect communication, or mixed with a certain type of cells for mixed cell co-culture, only It can form a co-culture structure system with partial layered structure or partial direct contact between vascular endothelial cells-astrocytes-neurons, so it is difficult to construct a fully independent layered structure and at the same time achieve a full chain of cells The direct contact neurovascular unit model of the direct communication link cannot effectively simulate the physiological functions of the blood-brain barrier, nutritional coupling, and electrophysiological coupling of the neurovascular unit in vivo.

Third, most of the current research on the neurovascular unit is limited to the improvement of neurovascular unit cell culture technology, and there is a lack of methodological research on the construction of independent layered direct contact connection structures between various cells of the neurovascular unit

[0008] In short, the existing in vitro co-culture cell models of the neurovascular unit have their own shortcomings, and it is difficult to fully and effectively simulate the physiological functions of the neurovascular unit in vivo

Images