Culture device for adherent cells, 2.5D beehive type culture system and method

A culture device and a technology for adherent cells, applied in the field of cell culture, can solve the problems of complex structure, low output, unsatisfactory, etc., and achieve the effect of increasing the number of cultures, avoiding damage, and increasing efficiency

Inactive Publication Date: 2021-06-18
湖南美柏生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing culture of adherent cells mainly has two forms: two-dimensional culture and three-dimensional culture. Market demand for exosomes; the three-dimensional culture method is to form cells and three-dimensional structure carriers (three-dimensional cell scaffolds or microsphere carriers) into a composite form of cells-three-dimensional carriers, and co-cultivate them in vitro to avoid cell contact inhibition during the culture process. The problem of reducing the amplification rate, but the three-dimensional structure carrier has a complex structure compared to the two-dimensional culture plate, the cost is high, and it is not easy to obtain the extracellular matrix through physical methods

Method used

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  • Culture device for adherent cells, 2.5D beehive type culture system and method
  • Culture device for adherent cells, 2.5D beehive type culture system and method
  • Culture device for adherent cells, 2.5D beehive type culture system and method

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Embodiment 1

[0063] Such as figure 1 As shown, the present invention provides a culture device for adherent cells, comprising a culture cylinder 1, a TC support 2 and a TC culture plate 3; the culture cylinder 1 is circular, and its material is other materials such as medium boron or high boron or quartz glass. High temperature and high pressure resistant material, after high temperature and high pressure sterilization, it is used to hold cell culture medium, TC support 2 and TC culture plate 3, and provides a sterile environment for cell culture. In this embodiment, the height of culture cylinder 1 is set to 15- 18cm, the inner diameter is set to 152-163mm, and the thickness is set to 3-7mm.

[0064] Wherein, the cultivation cylinder 1 has a cultivation cylinder cover 11, and the cultivation cylinder cover 11 is used to open and close the described cultivation cylinder 1, and is also used to communicate with the CO in the incubator. 2 Carry out gas exchange, specifically, the structure o...

Embodiment 2

[0069] This embodiment provides a 2.5D beehive culture system, the 2.5D beehive culture system includes cell culture medium and the culture device as described in the embodiment, the cell culture medium is filled in the culture tank, including adherent cell culture medium As well as macromolecular solutes, the adherent cell culture medium can be a conventional culture medium for culturing adherent cells, such as fetal bovine serum. The adherent cell culture medium in this embodiment not only includes fetal bovine serum, but also MEM medium and / or DMEM medium, antibiotics and glutamine, thereby forming double antibody culture fluid, macromolecule solute is one or more in carrageenan, xanthan gum and dextran sulfate in the present embodiment, present embodiment The cell culture medium can partially simulate the three-dimensional culture environment in vivo through the volume effect of macromolecular solutes, especially in the narrow gap of only 1.5-2mm between two adjacent TC cu...

Embodiment 3

[0071]This embodiment provides a method for cultivating adherent cells using the 2.5D beehive culture system described in Embodiment 2, comprising the following steps:

[0072] S1. Immerse each TC culture plate in an inoculation container with cell culture medium, such as an inoculation tray set in a biological safety cabinet, wherein the cell culture medium includes adherent cell culture fluid and macromolecular solute, and the macromolecular solute is 0.05 mg xanthan gum per mL, adherent cell culture medium includes 85ml of a-MEM medium and DMEM medium mixed in equal volume, 0.8ml of antibiotics, 8ml of fetal bovine serum, and 0.8ml of glutamine;

[0073] S2. Carry out the first cell inoculation on the cell culture surface provided with the support part on the TC culture plate, and cultivate for 2 hours; then turn over the TC culture plate, and perform the second cell inoculation on another cell culture surface , cultivated for 2 hours, and obtained the pretreated TC culture...

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Abstract

The invention discloses a culture device for adherent cells, a 2.5D beehive type culture system and method. The culture device comprises a culture jar, a TC bracket and TC culture plates, wherein the TC bracket is arranged in the culture jar and is provided with a plurality of fixing parts which are arranged at different heights, and the fixing parts are used for mounting a plurality of TC culture plates in a layered manner; and the two surfaces of each TC culture plate are cell culture surfaces, and the TC culture plates are detachably arranged on the fixing parts with different heights respectively to form multiple layers of cell culture surfaces. The 2.5D beehive type culture system comprises the culture device and a mixture of an adherent cell culture solution and a macromolecular solute. According to the method, the mixture is injected into the 2.5D beehive type culture system for culture. The culture device provided by the invention is favorable for increasing the culture area of unit volume; the provided 2.5D beehive type culture system can simulate a 3D culture environment; and the method provided by the invention can be used for promoting cells to secrete more extracellular matrix proteins.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a culture device for adherent cells, a 2.5D beehive culture system and a method. Background technique [0002] Cell culture is an essential process in bioengineering technology, which can transform a single cell into a simple single cell or rarely differentiated multi-cell through mass culture. A large number of cells or their metabolites can be obtained by cell culture. Since most biological products are obtained from cells, cell culture technology is the core and basic technology in biotechnology. Cell culture work has been widely used in various fields such as biology, medicine, and new drug research and development, and it plays a vital role in the entire life science research. [0003] Adherent cells must be attached to the surface of the support during the growth process to climb out of primary cells faster and better, such as smooth muscle cells, umbilical cord mese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/04C12M3/00
CPCC12M23/04C12M23/20C12M23/38C12M23/48C12M25/06
Inventor 肖锷
Owner 湖南美柏生物医药有限公司
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