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Separating material and method for collecting cell or the like using the same

a technology of material and cell, applied in the field of separation material and method for collecting cell or the like using the same, to achieve the effects of convenient and efficient harvesting method, superior film forming capability, and convenient and low cost production method

Inactive Publication Date: 2012-06-21
KANEKA CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0059]An object of the present invention is to provide a novel material having superior film forming capability, which is expected to show various physiological functions such as biocompatibility, blood compatibility, affinity for cell or protein, and the like, and a convenient and low cost production method thereof, provide a physiological material containing the material and an isolation material using the same, and to provide a convenient and efficient method of harvesting a cell such as monocyte and the like or a protein as well as a convenient and efficient method of preparing a dendritic cell, using the isolation material.Means of Solving the Problems
[0060]The present inventors have conducted intensive studies in an attempt to solve the above-mentioned problems and found that a cell such as monocyte and the like or a protein can be isolated conveniently and efficiently by the use of an isolation material containing, on at least a part of the surface, a substance having a sugar structure, and that a dendritic cell can be prepared conveniently and efficiently from a monocyte.
[0061]Furthermore, the present inventors have also found that, as the above-mentioned substance having a sugar structure, a sugar chain-containing polymer having a particular structure defined below particularly has superior film forming capability, is expected to have various physiological functions such as biocompatibility, blood compatibility, affinity for a cell or a protein and the like, and is useful for harvesting a cell such as monocyte and the like or a protein, and that the sugar chain-containing polymer can be produced by a convenient method at a low cost.

Problems solved by technology

However, the method of isolating monocytes by the density-gradient centrifugation method is associated with many problems yet to be solved, such as a safety problem of the liquid to be used for density gradient (cytotoxicity and the like), an operability problem because of a long time required for centrifugation, washing operation and the like, much loss of cells, an open system operation allowing easy contamination and the like, isolation efficiency (insufficient removal of non-adhesive cells, contamination with lymphocytes) and the like.
However, this method requires complicated operation, and the recovery rate and purity of the isolated monocytes are not sufficient.
The isolation method using magnetic beads with antibody can isolate monocytes at a comparatively high purity, but the cell treatment requires a long time, and also, is expensive.
This method affords high purity monocytes, but the apparatus is expensive, and requires a skillful operation technique.
In addition, the cell isolation problematically requires a long time.
In addition, the property is not sufficient because the monocyte recovery rate is about 50% and the purity is about 40%.
However, such method cannot isolate lectin sufficiently, and high purity lectin cannot be obtained problematically.
Nevertheless, since this method requires chemically binding a sugar ligand onto the carrier surface, the sugar density is not sufficient, lectin is not sufficiently isolated, and high purity lectin cannot be obtained problematically.
On the other hand, this may result in the degradation of the film forming capability, mechanical intensity and the like that skeleton polymers inherently show.
However, since the production step for introducing a sugar chain into the side chain requires complicated protection and deprotection operations, the production cost may problematically increase.patent document 1: JP-A-63-238105patent document 2: JP-A-63-68603patent document 3: JP-A-7-304788patent document 4: JP-A-8-253495patent document 5: JP-A-8-319317 (JP-B-3053764)patent document 6: JP-A-8-253495patent document 7: JP-A-2002-88094patent document 8: JP-A-2005-112987patent document 9: JP-A-5-140294patent document 10: JP-A-5-140213patent document 11: JP-A-2002-302511patent document 12: JP-A-5-178986patent document 13: JP-A-8-337566patent document 14: JP-A-9-227600patent document 15: JP-A-11-60603patent document 16: JP-A-2003-73397patent document 17: JP-A-11-71391patent document 18: WO98 / 32840patent document 19: JP-A-9-75076 (JP-B-3812909)patent document 20: JP-A-2004-129550patent document 21: JP-A-62-201641patent document 22: JP-B-2660175patent document 23: JP-A-10-504287patent document 24: JP-B-3711356patent document 25: JP-A-63-68603patent document 26: JP-B-1995189patent document 27: JP-A-11-315091patent document 28: JP-A-2004-526691patent document 29: JP-A-2005-15451non-patent document 1: Design and Physiology of Sugar Chain Molecule, 2001, Japan Scientific Societies Pressnon-patent document 2: Method of Studying Physiologically Active Sugar Chain, 1999, Japan Scientific Societies Pressnon-patent document 3: Carbohydrate Engineering and Production Technique, 1993, Science Forum Inc.non-patent document 4: Proc. Natl. Acad. Sci., vol.

Method used

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  • Separating material and method for collecting cell or the like using the same
  • Separating material and method for collecting cell or the like using the same
  • Separating material and method for collecting cell or the like using the same

Examples

Experimental program
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Effect test

example a

Harvesting Method of Monocyte and Preparation Method of Dendritic Cell

[0240]The present invention is explained in more detail by referring to the following Examples, which are not to be construed as limitative.

example a1

(1) Preparation of Cell Isolation Material

[0241]A mannobiose-bound polystyrene derivative (Poly[N-p-vinylbenzyl-O-β-D-mannopyranosyl-(1→4)-D-mannamide], manufactured by SEIKAGAKU CORPORATION) was dissolved in distilled water at a concentration of 0.1% (W / V). The solution (1 ml) was added to a commercially available 1 ml polystyrene petri dish (diameter 35 mm), and left standing at room temperature for about 1 hr. Then, the supernatant polymer was removed, and saline (1 ml) was added. Excess mannobiose-bound polystyrene derivative was washed, and the supernatant was removed. This operation was repeated 3 times to give a mannobiose-bound polystyrene derivative coated petri dish.

(2) Isolation of Monocyte

[0242]A winged injection needle was punctured into an upper arm of a healthy subject, and the blood (about 7.5 ml) was drawn in a lymphocyte isolation tube (vacutainer tube, manufactured by Becton, Dickinson and Company). After blood sampling, 3.8% citric acid solution was added to the ...

example a2

[0246]In the same manner as in Example A1 except that a lactose-bound polystyrene derivative (nonreducing terminal sugar is galactose, manufactured by SEIKAGAKU CORPORATION) was used instead of the mannobiose-bound polystyrene derivative, the recovery rate and purity of the monocyte were measured. As a result, the recovery rate of monocyte was 81% and the purity thereof was 73%, and the monocyte was found to have been selectively isolated and collected.

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Abstract

The present invention provides a novel material useful for selectively isolating a cell such as monocyte and the like or a protein from a body fluid and a production method thereof, a physiological material using the material and an isolation material using the physiological material, as well as a method of harvesting a cell such as monocyte and the like using the isolation material, a method of harvesting a protein and a method of preparing a dendritic cell.

Description

CROSS-REFERENCE[0001]This application is a divisional of U.S. application Ser. No. 12 / 302,881, filed Jun. 9, 2009, which is a National Stage application of PCT / JP2007 / 060721 filed May 25, 2001.TECHNICAL FIELD[0002]The present invention relates to a novel material useful for selectively isolating a cell such as monocyte and the like from a body fluid or a protein contained in an animal or plant and a production method thereof, a physiological material using the material and an isolation material using the same, as well as a method of harvesting a cell such as monocyte and the like, a method of harvesting a protein by the use of the isolation material, and a method of preparing a dendritic cell.BACKGROUND ART[0003]In recent years, it has been clarified that sugar chain present in the cell surface is deeply involved in various vital phenomena (cell recognition, transduction of information, cell adhesion, differentiation and proliferation, canceration, virus infection, blood coagulation...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N11/10C12N5/0784C12N5/0786
CPCC07K1/22C12N5/0645C12N5/0639C07K14/42
Inventor AWAJI, HIROSHIKUMAR, ASHUTOSHKOBAYASHI, AKIRAIMAI, NAOHIRO
Owner KANEKA CORP
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