Low-intensity pulse ultrasound-assisted rotary tubular cell/tissue culture system

A technology of pulsed ultrasound and tissue culture, applied in the field of biomedical tissue engineering, can solve the problems of inability to supply oxygen online, high price, high temperature resistance of the exchange membrane, etc., achieve good proliferation and differentiation, promote three-dimensional growth, and facilitate repeated use.

Inactive Publication Date: 2010-12-15
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0014] The purpose of the present invention is based on the inability of online oxygen supply and liquid supply in the existing cell culture device, the inability to accurately control the oxygen supply concentration, the exchange membrane is not resistant to high temperature, it is not convenient for disinfection, and no auxiliary means are used to stimulate the proliferation of cells / tissues , Differentiation, it is difficult to provide a good growth microenvironment, and the price is expensive. Provide a rotating tube cell / tissue culture system assisted by Lipus

Method used

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  • Low-intensity pulse ultrasound-assisted rotary tubular cell/tissue culture system
  • Low-intensity pulse ultrasound-assisted rotary tubular cell/tissue culture system
  • Low-intensity pulse ultrasound-assisted rotary tubular cell/tissue culture system

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Embodiment 1

[0045] The bone marrow mesenchymal stem cells (Bone Marrow Stem Cells, MSCs) cultured in the present invention are differentiated and proliferated into bone cells. Specific steps are as follows:

[0046] Mix the MSCs with the culture medium (where DMEM and serum-free DMEM are mixed at a volume ratio of 1:1) into the cell culture chamber, start the culture system, adjust the rotation speed to a suitable speed, and adjust the Lipus parameter to the frequency of 1.5MHz and the intensity of 30mW. / cm 2 , with a repetition rate of 1 kHz and a pulse width of 200 μs, at a temperature of 37 °C to start the rotational stimulation culture. The culture liquid is regularly supplied to the culture chamber through the online liquid supply system, the gas is supplied to the gas chamber in real time through the online gas supply system, and the oxygen concentration is controlled within an appropriate range through the oxygen concentration sensing system. After culturing for 48 h, the osteoc...

Embodiment 2

[0048] The present invention is used for joint culture of MSCs and "collagen / nano Tri-calcium Phosphate / Hyaluronic Acid" (Col / nano-TCP / HA) osteochondral scaffold material to construct tissue engineering cartilage. Specific steps are as follows:

[0049] The Col / nano-TCP / HA biomaterial was trimmed to a diameter of 5 mm and a thickness of 3 mm, and the third-generation MSCs were seeded into the scaffold material at a density of 1 × 106 / ml, and then mixed with the culture medium (wherein DMEM and serum-free DMEM were 1:1). 1 ratio mixing) and add it to the cell culture chamber, start the culture system, adjust the rotation speed to a suitable speed, and adjust the Lipus parameter to the frequency of 1.5MHz and the intensity of 2mW / cm 2 , with a repetition rate of 1 kHz and a pulse width of 200 μs, at a temperature of 37 °C to start the rotational stimulation culture. The culture liquid is regularly supplied to the culture chamber through the online liquid supply system, the gas ...

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Abstract

The invention discloses a low-intensity pulse ultrasound-assisted rotary tubular cell / tissue culture system. The Lipus-assisted rotary tubular cell / tissue culture system comprises a culture main body provided with a rotating shaft, wherein an air chamber is sheathed outside the culture main body, the rotating shaft is arranged at both ends of the air chamber, one end of the culture main body is provided with an Lipus ultrasound instrument, the ultrasound transmitting probe of the Lipus ultrasound instrument is inserted into the culture main body, and the inner wall of the air chamber is provided with an oxygen concentration controlling part. The culture system can realize on-line oxygen supply and liquid supply, thereby being beneficial to the transport of oxygen, carbon dioxide and metabolin in the system and being capable of accurately controlling the oxygen concentration. By using the Lipus ultrasound instrument as an auxiliary means, the invention creates a favorable microenvironment for the proliferation or differentiation of cells or tissues, ultimately simulates the micro-gravity environment for carrying out three-dimensional dynamic culture, and can realize functions of oriented amplification and induced differentiation.

Description

technical field [0001] The invention relates to the technical field of biomedical tissue engineering, in particular to a rotating tubular cell / tissue culture system assisted by low-intensity pulsed ultrasonic waves. Background technique [0002] The National Natural Science Foundation of the United States (NSF) proposed the concept of "tissue engineering" in an academic symposium held in August 1987, and clearly defined its connotation in 1988 as: "the principles of applied engineering science and life science and methods to elucidate the function-structure relationships of mammalian tissues and to develop active artificial substitutes to restore, maintain, or enhance tissue function." The key lies in the in vitro culture of mammalian cells to form living tissues or various biological substitutes with specific functions and apply them clinically. Tissue engineering involves many scientific issues, and the three-dimensional cell / tissue culture system is one of its important ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/42
CPCC12M25/02C12M23/06C12M35/04C12M41/34C12M27/06
Inventor 林子洪吴少旭陈奕春郭晓升陈耿东陈晓敏黄楚辉廖威明
Owner SUN YAT SEN UNIV
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