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Laser prepn process of directionally arranged nanometer channel regulating cell

A technology of directional arrangement and cells, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of unsuitable cell culture, high production cost, graphic defects, etc., and achieve the effect of convenient observation, low cost and simplified method

Inactive Publication Date: 2005-07-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Simply using grooved silicon wafers or quartz as a culture substrate has high production costs and low efficiency, and silicon wafers or quartz are not suitable for normal cell culture; using polymer molding to produce grooves, the temperature will increase during the molding process. Deformation of other parts of the polymer surface, and pattern defects or deformation during the demolding process, this phenomenon is particularly obvious for small-sized grooves

Method used

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  • Laser prepn process of directionally arranged nanometer channel regulating cell
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  • Laser prepn process of directionally arranged nanometer channel regulating cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Nanogrooves regulate directional growth and alignment of rat glioma (C6) cells.

[0030] 1. Preparation of nano-grooves on the surface of PS polymer: select the Nd:YAG laser 4 frequency doubling device, so that the wavelength of the laser output laser is 266nm; at the same time, use the mirror group of 266nm wavelength to make the laser reach the surface of the mobile platform vertically. Fix the clean PS Petri dish (35mm) on a support on an X-Y mobile operating platform. Turn on the laser, and after 40 minutes of warm-up, the laser will generate and output stably. The pulse width of the laser is 6ns and the frequency is 10Hz. Use an energy meter to detect and adjust the laser energy so that the energy reaching the surface of the sample is 30mJ / cm 2; At the same time, adjust the incident angle of the laser to be 20°, and the incident light is S polarized light. Start the control program of the mobile platform in the computer to move the platform accurately in the X-...

Embodiment 2

[0035] Nanogrooves regulate the directional growth of bone marrow stromal stem cells (MSCs) induced to differentiate into osteoblasts.

[0036] 1. Fabrication of nano grooves: the energy of the laser is 50mJ / cm 2 , adjust the incident angle of the laser to 30°, the moving speed of the platform in the X direction is 0.015mm / s, and the moving speed in the Y direction is 30mm / s. Others are the same as example 1.

[0037] 2. Soak the culture substrate in 75% medical alcohol for 24 hours for disinfection, then take it out and place it on a sterile operating table to dry.

[0038] 3. MSCs cell culture: the composition of the culture medium for inducing bone marrow stromal stem cells to differentiate into osteoblasts: containing 100nmol / L dexamethasone (Sigma), 50μg / ml vitamin C (Sigma, cultured pure), 10mmol / Lβ-glycerol phosphate , High Glucose DMEM (Sigma), 15% Fetal Bovine Serum.

[0039] A 4-month-old New Zealand white rabbit was anesthetized by intravenous injection of 1% pen...

Embodiment 3

[0041] Using changes in the orientation of nanogrooves to fabricate cell patterns

[0042] 1. According to the characteristic that nanogrooves can control the growth and arrangement of cells in a certain direction, nanogrooves with different directions are prepared on the same substrate, so as to regulate cell orientation and form cell patterning in different orientation directions. .

[0043] 2. Cell culture: C6 cells were selected as the cells, and the culture method was the same as in Example 1. When the direction of the nanogroove changes, the direction of the cell pattern changes accordingly. exist Figure 5 In A and B, as a comparison, the lower half of the figure is the region without nanogrooves, and the upper half is the region with nanogrooves. Figure 5 In A, the direction of the nanogroove is perpendicular to the untreated area, and the direction of the cell pattern is also perpendicular to the unoriented cells; when the angle between the direction of the nanogr...

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Abstract

The common medical polymer material to be used as cell culture substrate is treated through laser scanning to form micron / nanometer or submicon / nanometer channels before sterilization. Anchorage dependent cell in condition after trypsinization is inoculated homogeneously to the surface of the substrate for culture. Owing to the regulation and control of the surface channels, cell will grow and orient in the channel direction. The present invention has simple technological process and easy operation, and is suitable for the basic research in cell biology, making of biological sensor and functional device, research of medical implant, tissue engineering, etc.

Description

technical field [0001] The invention relates to a method for regulating the directional growth and arrangement of cells, in particular to a method for preparing nano-grooves on the surface of a cell culture substrate using laser LIPS (laser induced periodic surface structures) technology, and then for cell culture induction and control of cell directional growth The method of arrangement and alignment belongs to the category of biomaterials, and can be used for research in the fields of cell biology, implants and tissue engineering, as well as for the development of biosensors. Background technique [0002] The development of organ and tissue repair and transplantation, cell sensor, drug development, biochip, artificial neural network and life mechanism research in tissue engineering requires directional control of cell growth and arrangement. As the smallest unit of living organisms, cells can perform almost all functions of living organisms, such as metabolism, heredity, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
Inventor 朱邦尚路庆华陈万涛王宗光胡钧
Owner SHANGHAI JIAO TONG UNIV
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