Human blood cerebrospinal fluid barrier model as well as preparation method and application thereof

A cerebrospinal fluid and barrier technology, which is applied in the field of human blood and cerebrospinal fluid barrier model and its preparation, can solve the problems of simple model structure, limited experimental research, and difficulty in reproducing the physiological function of BCSFB, and achieves high repeatability, good compatibility, and operation. simple effect

Pending Publication Date: 2022-07-15
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the large differences between animal models and real human physiological conditions, and because the anatomical location of BCSFB is located deep in the brain and difficult to observe, its preclinical experimental research is limited; and in vitro models are mainly constructed using two-dimensional cells, and the model structure is too simple , it is difficult to reproduce the physiological function of BCSFB

Method used

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  • Human blood cerebrospinal fluid barrier model as well as preparation method and application thereof
  • Human blood cerebrospinal fluid barrier model as well as preparation method and application thereof
  • Human blood cerebrospinal fluid barrier model as well as preparation method and application thereof

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preparation example Construction

[0038] According to a typical embodiment of the present invention, a method for preparing a human blood and cerebrospinal fluid barrier model is provided, the method comprising:

[0039]Step S1, preparing a microfluidic chip with a biocompatible material by an inversion method, and the microfluidic chip contains an upper channel and a lower channel arranged adjacently;

[0040] The biocompatible material includes polyethylene terephthalate PDMS prepolymer; or type I collagen, matrigel, GelMA, sodium alginate, fibrinogen, HAMA, HA-CA, silk fibroin, chitosan and at least one of gelatin.

[0041] The step S1 specifically includes:

[0042] Step S101, obtaining a male mold with an adjacent upper channel and a lower channel;

[0043] The length, width and height of the upper channel and the lower channel are respectively 9000-11000 μm, 800-1200 μm and 20-200 μm.

[0044] If the height of the channel is less than 20μm, the internal hydraulic impedance is too large, the sample loa...

Embodiment 1

[0066] Embodiment 1. A human blood and cerebrospinal fluid barrier model and its preparation method

[0067] (1) The microfluidic chip is fabricated by a standard soft lithography flip-molding method. First, a silicon master mold etched with a micro-channel structure is fabricated by soft lithography. Secondly, the upper and lower layers of the chip are fabricated by a reverse molding method using PDMS. Subsequently, the chip was thoroughly dusted with 3M tape, and a piece of porous polyethylene terephthalate (PET) film (film thickness 7 μm, pore diameter 0.4 μm, pore density 0.4 μm) was flatly placed between the upper and lower channels under a stereo microscope. 4×10 6 / cm 2 ), align the upper and lower channels, and use a clamp made of poly-methyl methacrylate (PMMA) for fixed assembly. Finally, fully sterilize the assembled chips for later use;

[0068] (2) The upper channel of the chip was coated with Matrigel 80 μg / mL, and the lower channel was coated with Laminin (...

Embodiment 2

[0071] In the embodiment of the present invention, the length of the chip channel is increased by 20000 μm, and other structures and steps are the same as those in Embodiment 1.

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Abstract

The invention discloses a human blood cerebrospinal fluid barrier model as well as a preparation method and application thereof, the method comprises the following steps: a microfluidic chip is prepared from a biocompatible material through a reverse molding method, and the microfluidic chip contains an upper channel and a lower channel which are adjacently arranged; arranging a porous film between the upper channel and the lower channel of the micro-fluidic chip, and fixing the micro-fluidic chip by using a clamp to obtain a culture device; placing the culture device in a matrix material, so that the matrix material coats the peripheries of the upper channel and the lower channel; and injecting a human venation plexus epithelial cell suspension into the upper channel, injecting a human brain microvascular endothelial cell suspension into the lower channel, and after cells are attached, applying fluid stimulation to the upper channel and the lower channel and culturing to obtain the human blood cerebrospinal fluid barrier model. The model is high in bionic degree, easy to detect and suitable for optical imaging instruments such as confocal microscopes and the like.

Description

technical field [0001] The invention relates to the technical field of tissue engineering and organ chips, in particular to a human blood and cerebrospinal fluid barrier model and a preparation method and application thereof. Background technique [0002] BCSFB is located in the lateral ventricle, the third ventricle and the fourth ventricle, and is composed of choroid plexus epithelial cells (CPECs) that are tightly connected to each other and have polarity, and brain microvascular endothelial cells (brain microvascular endothelial cells) with fenestrated structures. cells, BMECs). In addition to the physical barrier function, choroid plexus epithelial cells are the main site of CSF secretion and reabsorption, and can secrete a variety of cytokines to maintain nervous system stability. Studies have found that the use of human cerebrospinal fluid is closely related to neurogenesis, gliogenesis, synapse formation, neurite outgrowth and other neurodevelopmental processes, ind...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/02B01L3/00
CPCC12N5/069C12N5/0625G01N33/5005B01L3/5027C12N2502/00C12N2503/02C12N2521/00C12N2533/90C12N2533/52G01N2500/10B01L2200/10B01L2300/12
Inventor 陈璞彭勉周莹徐放
Owner WUHAN UNIV
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