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Stem cell globe cutting and collecting device and stem cell globe cutting and separating passage method

A stem cell ball and collection device technology, applied in the field of cell culture, can solve the problems of long operation time, cell damage, uneven size of neurospheres, etc., and achieve the effect of controllable quality and stable characteristics

Active Publication Date: 2015-12-23
ZHEJIANG ORIGIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention provides a cutting and collecting device for stem cell balls in order to solve the problems of neural stem cells prone to apoptosis, mutation, cell damage, neural stem cell differentiation, long operation time and uneven size of neurospheres caused by neural stem cell passaging process

Method used

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  • Stem cell globe cutting and collecting device and stem cell globe cutting and separating passage method
  • Stem cell globe cutting and collecting device and stem cell globe cutting and separating passage method
  • Stem cell globe cutting and collecting device and stem cell globe cutting and separating passage method

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Such as figure 1 and figure 2 A cutting and collecting device for stem cell spheres is shown, which consists of a stem cell sphere cutter 6, an air buffer bottle 2, a three-way pipe 4, a culture bottle 5, a stem cell sphere collection bottle 3, a small stem cell sphere collection bottle 9 after cutting and a It consists of an air pump 1 with positive pressure and negative pressure two-way air suction functions.

[0030] The stem cell sphere cutter and the stem cell sphere collection bottle are connected by a pipeline. The stem cell sphere cutter has a tube body, and two screens 7 arranged at intervals are fixed in the tube body. The sieve is a stainless steel sieve, the apertures of the two sieves are 350 μm and 104 μm respectively, the 350 μm sieve is close to the stem cell sphere collection bottle, and the distance between the two sieves is 20 mm. The outlet end of the stem cell sphere cutter 6 is provided with a constricted connecting pipe 8 .

[0031] The air pu...

Embodiment 2

[0036] Such as Figure 4 The device for cutting and collecting stem cell spheres includes a syringe 10 and a stem cell sphere cutter 6 connected to the syringe. The stem cell sphere cutter has a tube body in which two screens 7 are fixed at intervals. The volume of the syringe 10 is 100-200ml, and the outlet end of the stem cell sphere cutter 6 is provided with a constricted connection tube 8 . The screens are stainless steel screens, the apertures of the two screens are 350 μm and 104 μm respectively, and the screen with smaller aperture is close to the small stem cell sphere collection bottle 9 after cutting. The distance between the two screens is 20mm. The device is suitable for laboratory or small-scale cultivation of neural stem cells.

[0037] First draw the syringe into the 100ml of culture medium containing stem cell spheres to be cut, and use the thrust of the artificial hand to quickly pass the 100ml of culture medium containing stem cell spheres through the stem ...

Embodiment 3

[0039] A method for cutting, separating and subcultureing neural stem cell spheres, the method comprising the steps of:

[0040] a. Use the stem cell ball cutting and collecting device described in Example 1 to cut and separate the stem cell balls: use the negative pressure of the air pump to suck the culture medium containing stem cells into the stem cell ball collection bottle, and then use the positive pressure of the air pump to make the stem cell balls Accompanied by the rapid passage of the liquid through the stem cell ball cutter, blunt cutting into small stem cell clumps, and then into the culture bottle, the cell culture medium is preset in the culture bottle, Figure 5 Neural stem cell spheres before cutting, Figure 6 Cut into small pieces for neural stem cell spheres, Figure 7 Individual neural stem cells and small clumps for microscopy at 400X magnification.

[0041] b. The cell culture fluid containing small stem cell clusters obtained in step a is cultured in...

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Abstract

In order to solve the problems of cell apoptosis, mutation, cell damage, and neural stem cell differentiation of neural stem cells, long operating time, uneven sizes of cell globes and the like which are caused in the passage process of neural stem cells, the invention provides a stem cell globe cutting and collecting device. The stem cell globe cutting and collecting device comprises a stem cell globe cutter, a stem cell globe collecting bottle, and an air pump provided with positive-pressure and negative-pressure two-way air extracting functions, wherein stem cell globe cutter is connected with the stem cell globe collecting bottle through a pipeline, the stem cell globe cutter is provided with a pipe, and two sieves arranged at an interval are fixed in the pipe. With the adoption of the device, and a stem cell globe cutting and separating passage method, the technical problem of large possibilities of differentiation and death existing in the in-vitro culture of the neural stem cells is solved, the purposes of long-term culture and amplification of the human neural stem cells are achieved, and in addition, the properties of the human neural stem cells are stable, and the quality of the human neural stem cells is controllable.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a device for cutting and collecting stem cell spheres and a method for cutting, separating and passing stem cell spheres. Background technique [0002] When culturing neural stem cells or neural precursor cells, tumor stem cells, and embryonic stem cells, stem cell clumps or stem cell spheres often appear. In long-term culture, trypsin or trypsin plus EDTA is used to separate and pass on cell spheres. Due to the long-term use of chemical separation of these enzymes, the cell membrane receptors are damaged to varying degrees, which seriously affects the lifespan of stem cells in long-term culture. The occurrence and development of nervous system diseases are caused by the decrease in the number of nerve cells or the change in function. Related neurodegenerative diseases can be treated by transplanting neural stem cells. Embryonic stem cells not only play a very ...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12N5/0797
CPCC12M33/04C12M33/12C12M33/14C12N5/0623
Inventor 陈世明章瑾
Owner ZHEJIANG ORIGIN BIOTECH
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