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Three-dimensional perfusion type cell culture device and application thereof

A cell culture and perfusion technology, applied in the field of cell culture equipment or devices, can solve the problems of high medical expenses, high mortality rate of acute and chronic renal failure, short dialysis life, etc. long effect

Active Publication Date: 2015-07-15
康珞生物科技(武汉)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high medical costs, short dialysis lifespan (typically only a few years) and scarcity of donor organs contribute to the high mortality rate in acute and chronic renal failure

Method used

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  • Three-dimensional perfusion type cell culture device and application thereof
  • Three-dimensional perfusion type cell culture device and application thereof
  • Three-dimensional perfusion type cell culture device and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] A three-dimensional perfusion cell culture device, including a box body 1, a cover body 2 and an elastic clip 3, the cover body 2 is arranged above the box body 1 and is movably connected to the box body 1 through a connector 4, the top surface of the box body 2 A first circular groove 5-1 is provided, and a liquid inlet channel 6-1 communicating with the first circular groove 5-1 is horizontally arranged on the side wall of the box body 1; The other side wall is horizontally provided with a liquid outlet channel 6-2 communicating with the first circular groove 5-1; the axis of the liquid inlet channel is staggered with the axis of the liquid outlet channel; The second circular groove 5-2 with the same radius of the circular groove 5-1; the first circular groove 5-1 is coaxial with the second circular groove 5-2, and the first circular groove 5 The bottom surface of -1 is provided with the first fiber layer 9-1, and the bottom surface of the second circular groove 5-2 i...

Embodiment 2

[0049] isolated cells

[0050] Isolation of kidneys from neonatal rabbits:

[0051] 1. Preparation: Surgical scissors, tweezers, etc. are pre-sterilized, and a 100ml small beaker is filled with IMDM culture solution without phenol red. The culture solution is purchased from Gibco, product number 21056#; Hazineethanesulfonic acid (abbreviated as HEPES) to a final concentration of 5%;

[0052] 2. Take the newborn rabbit baby within 2 days from the animal room. In the fume hood, moisten a thick cotton pad with ether and place it in a round glass dish, place the newborn rabbit in the round glass dish containing ether for 1-2 minutes to anesthetize;

[0053] 3. Place the anesthetized rabbit baby on a thick white cotton pad, cut off the artery in the neck, and make it dead;

[0054] 4. Rinse the rabbit abdomen with 70% ethanol;

[0055] 5. Cut open the lower abdomen of the baby rabbit, carefully take out two small kidneys the size of soybeans, put them in IMDM culture medium, an...

Embodiment 3

[0063] An application of a three-dimensional perfusion cell culture device in cell culture, the specific steps are as follows:

[0064] The utensils and tools used in this example have all been sterilized.

[0065] (1) Clip the polyester fiber (hereinafter referred to as vessel) in the culture device into a plate containing IMDM culture solution with tweezers, and soak it repeatedly to remove the air bubbles therein;

[0066] (2) Under the binocular magnifying glass, the embryonic membrane in the petri dish is unfolded with needle-nose tweezers, and placed on a circular vessel. After laying flat, add another round vessel to become a typical "sandwich" tissue explant sandwich culture, image 3 ;

[0067] (3) One end of the latex conduit is placed in the culture bottle (containing 500ml IHA working culture fluid), and is connected with the liquid inlet channel of the cell culture device through a high-precision peristaltic pump; the other latex conduit connects the liquid outl...

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Abstract

The invention discloses a three-dimensional perfusion type cell culture device and an application thereof. The three-dimensional perfusion type cell culture device comprises a box body, a cover body and an elastic clamp, wherein a first circular groove is formed in the top surface of the box body, and a second circular groove with the same radius as the first circular groove is formed in the bottom surface of the cover body. In the cell culture device disclosed by the invention, vessels consisting of polyester fibers are arranged on the bottom surface of the groove, and a cell or tissue explant is positioned between two layers of vessels to form 'sandwich' culture; the vessels consisting of polyester fibers are used for replacing a scaffold material, so that the dead angle volume in cell culture is minimized to better conform to the in-vivo physiological microenvironment. A culture solution inlet channel and a culture solution outlet channel are formed on the side wall of the box body of the culture device respectively, and a culture solution flows into the groove cavity through the solution inlet channel and flows out through the solution outlet channel so as to realize perfusion culture. By adopting the device disclosed by the invention, the embryonic stem cell / progenitor cell at the new renal cortex part is successfully differentiated into renal tubule tissues.

Description

technical field [0001] The invention belongs to the field of cell culture equipment or devices, and in particular relates to a three-dimensional perfusion cell culture device, and also relates to the use of a three-dimensional perfusion cell culture device. / A cell culture vessel for successful differentiation of progenitor cells into renal tubular tissue. Background technique [0002] Since Willhelm Roux isolated cells from chicken embryos in 1885 to establish in vitro cell culture for the first time, monolayer cell culture technology has a history of more than 100 years. For more than a century, monolayer cell culture has flourished. Especially in industrial fields such as pharmaceuticals or vaccine synthesis, products can be manufactured efficiently through rapid cell division. However, in the field of basic life science research, for the in vitro culture of cells, the focus is not only on their division and growth, but more importantly, whether they can maintain their...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/00
CPCC12M21/08C12M23/20C12M25/00C12M29/10
Inventor 胡康洪
Owner 康珞生物科技(武汉)有限公司
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