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Method for regulating distance between two species of cells

A cell and distance technology, applied in the field of materials and cells, can solve the problem of limited ability to regulate the distance between cells, achieve the effect of continuous controllable film thickness, low cost, and avoid materials and processes

Inactive Publication Date: 2017-06-20
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of transwell membranes to regulate the distance between two types of cells has the following problems: first, the ability to regulate the distance between cells is limited, only 10 μm and 1 mm; second, cells cultured on culture plates or gel membranes are in 2D growth Way

Method used

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  • Method for regulating distance between two species of cells
  • Method for regulating distance between two species of cells
  • Method for regulating distance between two species of cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: A method for regulating the distance between hepatocytes and fibroblasts

[0025] The 15mg / mL sodium alginate solution and the 3.6mg / ml collagen solution were mixed evenly at a volume ratio of 1:2, so that the final concentrations of the sodium alginate and collagen solutions were 5mg / mL and 2.4mg / mL, respectively. Then mix the human fibroblasts with the mixed sodium alginate and collagen solution, and adjust the cell density to 2×10 6 cells / mL. 300 μl of the cell suspension was slowly added to a circular glass slide with a diameter of 3 cm placed in a cell culture plate, and placed in a cell culture incubator for 2 hours to make the collagen gel. Add 1 mL of 100mM CaCl 2 The solution was added to the cell culture plate, calcified for 20min, sodium alginate and Ca 2+ Chelation forms calcium alginate gel, and finally collagen / calcium alginate gel is obtained. Then, add 1.5mg / mL chitosan (molecular weight 65kDa, degree of deacetylation 90%) solution, by con...

Embodiment 2

[0026] Embodiment 2: A kind of method for regulating the distance between human embryonic stem cells (hES) and fibroblast cells

[0027] First, arginine-glycine-aspartic acid (RGD) modified sodium alginate was prepared. Sodium alginate (molecular weight 430kDa, ratio of guluronic acid to mannuronic acid 1.5) was dissolved in 2-(N-morpholino)ethanesulfonic acid (MES) buffer (pH 6.5) containing 0.5M NaCl ), to obtain a 1% (W / V) sodium alginate solution. Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), N-hydroxysulfosuccinimide (sulfo-NHS) and RGD polypeptide, and stir at room temperature for 24 hours. The molar ratio of EDC to sodium alginate was 1:20, the molar ratio of EDC to sulfo-NHS was 2:1, and the mass ratio of RGD to sodium alginate was 1:1000. Then dialyzed and freeze-dried to obtain RGD-modified sodium alginate.

[0028] Afterwards, 5 mg of RGD-modified sodium alginate was weighed and dissolved in 100 mL of physiological saline to prepare a sodium alginate s...

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Abstract

After a species of cells and a polyanion (or polycation) polymer solution or( / and) main components of extracellular matrix are evenly mixed, the mixture is added on the surface of a round glass slide to prepare flat gel; then the flat gel is soaked in a polycation(or polyanion) polymer solution, the reaction time is controlled to obtain a polyelectrolyte hydrogel membrane with an adjustable thickness, then another species of cells is inoculated on the surface of the hydrogel, and thus the action distance between two species of cells can be adjusted by changing the thickness of the flat gel membrane.

Description

technical field [0001] The invention relates to the fields of cells, materials and the like, and is a method for regulating the distance between two types of cells. Background technique [0002] In vivo, the formation and function of tissues depend on the interaction between different types of cells. The interaction between cells is mediated through the mutual contact between cells and the exchange of soluble factors (paracrine effect). [Bhatia SN, Balis UJ, Yarmush ML, Toner M. Effect of cell-cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells. Faseb Journal. 1999; 13:1883-900.]. At present, more and more studies have found that in different time and space states, the interaction between cells presents different modes of action. When the distance is close, there are both cell-to-cell contact-mediated communication and paracrine effects between the two cells, so the function of the target cell [HuiEE, Bhatia SN. Mi...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12N5/10C12N5/0735
Inventor 于炜婷宋益哲王秀丽娄茹云郑会珍马小军
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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