JP2009254340A - Stainless steel cell cultivation apparatus and cell transfer container - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To settle the problems that stainless steel for cultivation apparatus and cell transfer containers has microscopic innumerable scratches, and thereby, pollution due to bacteria, virus, mycoplasma and the like, and cross pollution with other cells are caused, and pyrogen remains, wherein, in the cultivation apparatus, cells usable for regeneration medicine, such as ES cells, iPS cells and autologous skeletal myoblasts, and cells producing substances for raw materials for pharmaceuticals are cultivated. <P>SOLUTION: The surface on which no scratch even microscopically exists is obtained by a first surface treatment for making the stainless steel surface ≥0.1 μm but ≤1.0 μm ruggedness in a region of 250 μm length and width by a specific mechanical polishing method and then a second surface treatment for performing electrolytic polishing by using a specific electropolishing liquid. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a culture apparatus for culturing cells used for regenerative medicine, such as ES cells, iPS cells, autologous skeletal myoblasts, and cells that produce a raw material for pharmaceuticals, and a cell transfer container, particularly stainless steel. The present invention relates to a stainless steel cell culture device and a cell transfer container that reduce the adhesion of dirt, bacteria, and viruses by treating the surface, and improve the sterilization property by washing and chemicals.

In recent years, a technique for collecting a small amount of cells or tissues from each part of the human body, culturing them with an incubator, and using them for regenerative medicine has been developed. Devices for culturing cells have been developed. (Patent Document 1) However, culturing a plurality of types of cells in one facility inevitably means that other people's cells are present in the same space. Concerns about contamination of cells, bacteria, viruses, mycoplasma, etc. attached to them and cross contamination with other cells are concerned.
Therefore, every time a cell culturing operation is completed, chemical sterilization with ozone gas or autoclave sterilization with high-temperature steam is performed, and the stainless steel surface used in the apparatus is [Claim 1] or [Claim 2] or If the surface satisfies [Claim 3], drug sterilization and autoclave sterilization can be more effectively sterilized. In particular, there is almost no residual pyrogen which is a concern in autoclave sterilization.

In addition to regenerative medicine, a cell culture apparatus for culturing cells as raw materials for pharmaceuticals and the like has been developed to cultivate target cells in large quantities in a tank (Patent Document 2).
In the case of this equipment, there is concern about contamination of bacteria, viruses, mycoplasma, etc. from the wetted part of the tank (the part in contact with the cell culture solution) and cross-contamination with other cells. If the surface satisfies [Claim 1], [Claim 2], or [Claim 3], sterilization with drugs and autoclave sterilization can be performed more effectively, so that contamination of cultured cells can be effectively prevented.
Patent publication 2008-54690 Automatic cell culture device and method of use thereof. Patent publication 2008-43301 Cell culture method. Japanese Patent Application No. 2007-284361 Stainless equipment for doctors

An object of the present invention is to solve the above-mentioned problems and to obtain a cell culture device and a cell transfer container that effectively prevent contamination of cultured cells because drug sterilization and autoclave sterilization can be more effectively sterilized. .

In the present invention, the mechanical polishing by the triacact pyramid (Fig. 2) (Patent Document 3) manufactured by 3M Corporation is the optimum mechanical polishing, but electrolytic abrasive polishing, also known as electrolytic composite polishing, is sufficient if manufacturing costs are not an issue. The goal can be achieved. Further, if a slight performance deterioration is allowed, the conventional abrasive polishing + buff polishing can be used.
By performing electropolishing after these mechanical polishings, cell culture equipment and cell transfer containers with little contamination with bacteria, viruses, mycoplasma, etc., cross-contamination with other cells, and residual pyrogens are obtained.

The cell culture apparatus and the cell transfer container of the present invention have the effect that the stainless steel surface is less contaminated with bacteria, viruses, mycoplasma, etc., cross-contaminated with other cells, pyrogen remains, and is less rusting than the original material due to chromium concentration. As a result, it is possible to obtain an extremely high effect of cleaning and sterilization.

1) Examination of stainless steel material Steel type SUS316L Production method Vacuum double melting material, heat treatment method Bright annealing is best, but VOD material or AOD material can be produced if performance degradation is allowed.
2) Surface treatment before electropolishing.
Tri-Sact Pyramid or Electrolytic Abrasive Polishing, also known as Electrolytic Compound Polishing, manufactured by 3M, USA, which results in a surface with only uniform and shallow flaws without deep flaws. Can be obtained.
3) Electropolishing method.
Electrolytic polishing liquid is 85% phosphoric acid 60w%, 98% sulfuric acid 30w%
Additive ASLES EP2 (Okuno Pharmaceutical) 5w%
Pure water 5w% Temperature 60 ℃ ~ 80 ℃
Cathode electrode material SUS316L
Current density 1.2 A / in 2 to 2.4 A / in 2 Electrolytic polishing time 5 minutes to 10 minutes 4) Degreasing and cleaning before and after electrolytic polishing may be in a common sense range.

Hereinafter, one embodiment will be described with reference to the accompanying drawings, FIG. 1 is a container for transporting cultured cells,
It is sectional drawing of what SUS316L of thickness 1 and 0 mm, the inner and outer surface of a bright annealing material were grind | polished mechanically by 3M company TRIACT PYRAMID A-10, and also the inner and outer surface was electropolished.

It is sectional drawing of what SUS316L of thickness 1 of Example 1 and the bright-annealed material inner and outer surfaces were mechanically polished with 3M Corp. Trituract pyramid A-10, and the inner and outer surfaces were further electropolished. It is the surface photograph of a triacact pyramid.

Explanation of symbols

1 cultured cell transfer container.
Mechanical polishing and electrolytic polishing with a triacact pyramid are performed on the inner surface of the 1a cultured cell transfer container.
The outer surface of the 1b cultured cell transfer container is subjected to mechanical polishing and electrolytic polishing using a triacact pyramid.
2 Lid of cultured cell transfer container 2a Mechanical polishing and electrolytic polishing by triacact pyramid are performed on the inner surface of the lid of cultured cell transfer container.
3 packing 4 A catch clip that closes the lid.
5 handles.

Claims (7)

  The surface roughness is 0.03 μm or more and less than 0.3 μm in the vertical and horizontal 250 μm ranges, and the surface chromium concentration is 1.0 to 3.5 times that of iron. A cell culture apparatus and a cell transport container characterized by the above. Consists of SUS316L stainless steel,
After the first surface treatment in which the surface of the stainless steel is mechanically polished using an abrasive to make the surface irregularities 0.1 μm or more and 1.0 μm or less in a vertical and horizontal range of 250 μm, the second Mixed electrolytic polishing solution of phosphoric acid and sulfuric acid as the next surface treatment. Sodium nitrate / water mixed electrolytic polishing solution. A cell culture device characterized in that the chromium concentration on the surface is 1.0 times or more and 2.0 times or less that of iron by electropolishing using any one of a perchloric acid / ethanol mixed electropolishing liquid, and Cell transfer container. Consists of SUS316L stainless steel,
After the first surface treatment in which the surface of the stainless steel is mechanically polished using an abrasive to make the surface irregularities 0.1 μm or more and 1.0 μm or less in a vertical and horizontal range of 250 μm, the second Mixed electrolytic polishing solution of phosphoric acid and sulfuric acid as the next surface treatment. Electropolishing is performed using either sodium nitrate / water mixed electrolytic polishing liquid or perchloric acid / ethanol mixed electrolytic polishing liquid, and further immersed in nitric acid or citric acid so that the surface chromium concentration is 1.0 times that of iron. A cell culture device and a cell transfer container characterized by being 3.5 times or less. The cell culture device according to claim 1, 2 or 3, wherein the cell culture device is a tank type culture device. The cell culture device according to claim 1, 2, or 3, wherein the cell culture device is a multiple petri dish type automatic culture device. A cell culture device, wherein the abrasive according to claim 2 or 3 is a triazac pyramid. 4. The cell culture apparatus according to claim 2, wherein the mechanical polishing is electrolytic abrasive polishing, also known as electrolytic composite polishing.

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