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Cell sphere in-situ preparation method based on hydrogel microwell plate

An in-situ preparation and hydrogel technology, applied in the field of biomedicine, can solve problems such as low efficiency, easy loss of cells, and uneven density of planted cells, achieving broad application prospects, wide application range, and good repeatability

Active Publication Date: 2015-05-20
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently a lack of an effective method for constructing spherical micropores on the surface of the hydrogel; and, the current research is to artificially plant cells after the construction of microwell plates, which has low efficiency, uneven cell density, and the formation of spheres. Cells are easy to lose during the process

Method used

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  • Cell sphere in-situ preparation method based on hydrogel microwell plate
  • Cell sphere in-situ preparation method based on hydrogel microwell plate
  • Cell sphere in-situ preparation method based on hydrogel microwell plate

Examples

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Embodiment 1

[0059] A method for in situ preparation of cell spheroids based on a hydrogel microwell plate, comprising the following steps:

[0060] 1) The glass slide is subjected to hydrophobic treatment to obtain a hydrophobic substrate; the hydrophobic treatment is to soak the glass slide in a water-repellent agent and then dry it; the water-repellent agent is Zi Xilai self-cleaning agent;

[0061] 2) Digest the cultured MCF-7 cells, then mix with the first hydrogel sol to make the first hydrogel sol cell mixture; the cell concentration in the first hydrogel sol cell mixture is 1×10 6 a / mL;

[0062] Wherein, the first hydrogel sol is gelatin with a mass concentration of 3%;

[0063] 3) Use a pipette gun to deposit the first hydrogel sol-cell mixture in the form of 0.48 μL per droplet on the surface of the hydrophobic substrate prepared in step 1) to form hemispherical droplets, and stand at 4°C for 30 minutes , to cross-link and solidify hemispherical droplets to prepare cell-loaded ...

Embodiment 2

[0069] A method for in situ preparation of cell spheroids based on a hydrogel microwell plate, comprising the following steps:

[0070] 1) The polymethyl methacrylate substrate is subjected to hydrophobic treatment to obtain a hydrophobic substrate; the hydrophobic treatment is to paste a polytetrafluoroethylene film on the polymethyl methacrylate substrate to obtain a hydrophobic surface;

[0071] 2) Digest the cultured MCF-7 cells, then mix with the first hydrogel sol to make the first hydrogel sol cell mixture; the cell concentration in the first hydrogel sol cell mixture is 5×10 5 a / mL;

[0072] Wherein, the first hydrogel sol is methacrylic acylated gelatin with a mass concentration of 5%;

[0073] 3) Use a syringe to deposit the first hydrogel sol-cell mixture in the form of 0.23 μL per droplet on the surface of the hydrophobic substrate prepared in step 1) to form hemispherical droplets, and let it stand at 0°C for 30 minutes to make Hemispherical droplets are cross-l...

Embodiment 3

[0078] A method for in situ preparation of cell spheroids based on hydrogel microwell plates, comprising the following steps:

[0079] 1) The polymethyl methacrylate substrate is subjected to hydrophobic treatment to obtain a hydrophobic substrate; the hydrophobic treatment is performed to silanize the polymethyl methacrylate substrate to obtain a hydrophobic surface;

[0080] 2) digest the cultured cells, then mix with the first hydrogel sol to make the first hydrogel sol cell mixture; the cell concentration in the first hydrogel sol cell mixture is 0.8×10 7 a / mL;

[0081] Wherein, the first hydrogel sol is gelatin with a mass concentration of 5%;

[0082] 3) Use the cell printing platform to deposit the first hydrogel sol-cell mixture in the form of 0.48 μL per droplet on the surface of the hydrophobic substrate prepared in step 1) to form hemispherical droplets, and stand at 4°C for 10 minutes , to cross-link and solidify hemispherical droplets to prepare cell-loaded micr...

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Abstract

The invention discloses a cell sphere in-situ preparation method based on a hydrogel microwell plate and belongs to the technical field of biomedicines. The cell sphere in-situ preparation method comprises the following steps of firstly preparing a hydrophobic substrate and a cell-loaded microscale hydrogel template, preparing the hydrogel microwell plate in a reverse mold manner and carrying out cell culture in an incubator to form the cell sphere in situ. According to the method, the hydrogel microwell plate with spherical microwells is constructed by virtue of the cell-loaded microscale hydrogel as the template, the subsequent artificial cell inoculation is not needed, the cell density in the microwells is controllable and the method is easy to construct the size-controllable and regular-shape cell sphere. The method has the advantages of low cost, simple operation, good reproducibility, wide application range and has broad application prospects in fields of tissue engineering, regenerative medicine, drug screening and the like.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to a method for preparing cell spheres, in particular to an in-situ preparation method for cell spheres based on a hydrogel microporous plate. Background technique [0002] Cell spheres are a kind of multicellular aggregates. Compared with two-dimensional cell culture systems, cell sphere culture can better simulate the in vivo microenvironment and obtain response results that are closer to those in vivo. Therefore, in pathological mechanisms, drugs It has broad application prospects in screening and tissue engineering. There are many ways to construct cell spheroids, such as the microplate method based on the non-adherent surface of cells, the suspended droplet method based on gravity, the magnetic levitation method based on magnetic force, and the rotational culture method based on centrifugal force, etc. Among them, the microplate method has been widely used because of its simp...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/077C12N5/071C12N5/0775C12N5/0735C12N5/0797
Inventor 徐峰黄国友凌楷卢天健
Owner XI AN JIAOTONG UNIV
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