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Bioreactor for cell and tissue culture

A technology of bioreactor and cell culture, which is applied in the field of bioreactor to achieve the effect of reducing cost, improving water retention and increasing efficiency

Inactive Publication Date: 2008-12-31
DRUGMODE APS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although well known and widely used, currently available microgravity bioreactors have significant limitations:
However, none of these patents or published patent applications addresses the problem of dehydration or discloses microgravity bioreactors with small incubator volumes or with port closures of zero volume

Method used

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  • Bioreactor for cell and tissue culture
  • Bioreactor for cell and tissue culture
  • Bioreactor for cell and tissue culture

Examples

Experimental program
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Effect test

Embodiment approach

[0108]In a preferred embodiment, the semipermeable filters used in the present invention allow passage of molecules up to a certain molecular weight or size. Semi-permeable filters with defined pore sizes are known to those skilled in the art and are commercially available. In a preferred embodiment of the invention, the semi-permeable filter is permeable to molecules of a maximum predetermined molecular weight (eg 50 kDa, 100 kDa, 150 kDa, 200 kDa or 250 kDa). Alternatively, the permeability of a semi-permeable filter can be determined by the pore size therein. The pore size of the semi-permeable filter may be less than or equal to 0.5 μm, such as less than or equal to 0.3 μm, preferably less than or equal to 0.2 μm, more preferably less than or equal to 0.1 μm, and most preferably less than or equal to 0.05 μm. A wide variety of filters can be used. These filters may be made of materials selected from, but not limited to, polytetrafluoroethylene (PTFE), polyvinylidene fluo...

Embodiment 1

[0229] Example 1: Maintenance of the differentiated state of human hepatocytes in long-term culture: expression of cellular proteins

[0230] Specific proteins expressed by fully differentiated human livers and not normally found in hepatocytes grown under standard-plating culture conditions were found in cells cultured with the bioreactor of the present invention. Fully differentiated hepatocyte proteins were still found in culture after 302 days.

[0231] Human hepatocytes were grown in the bioreactor of the present invention for 302 days and then labeled with [35S]-methionine for 20 hours.

[0232] One or more cell cultures are grown as described above using a medium such as Eagles, DMEM or RPMI 1640 under normal culture conditions (eg 37°C for human cells). Depending on the specific cell culture considered, the medium needs to be changed every two or three days. Hepatocytes require fresh medium every two days. The medium was replaced by stopping the bioreactor, waiting ...

Embodiment 2

[0238] Example 2: Maintenance of the differentiated state of human hepatocytes in long-term culture: viability of the secretory mechanism

[0239] Hepatocytes cultured under microgravity conditions using the bioreactors of the present invention secrete specific proteins that are secreted by fully differentiated normal human mature hepatocytes and not normally secreted by hepatocytes grown under standard-plate culture conditions.

[0240] Proteins were precipitated from growth medium aliquots of the tissue culture of Example 1 and analyzed by two-dimensional gel electrophoresis and mass spectrometry. Proteins were identified by protein, accession number and protein description in Table 2. The number of isoforms identified correlates with gel staining spots that were positively identified as the protein of interest.

[0241] Table 2: Proteins found in bioreactor medium secreted by fully differentiated hepatocytes and not normally secreted by hepatocytes grown under standard-pla...

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Abstract

The present invention provides a bioreacto 1r for incubation of cell cultures, tissue biopsies, cell clusters, tissue-like structures,apted for rotation for use in microgravity conditions and equipped with an incubation cavity having a small internal fluid volume, generally less than 1 ml. To avoid problems associated with small volume incubation, the bioreactor may include a humidity chamber or other means of avoiding dehydration as well as substantially fluid-tight closures for access ports that avoid introduction of air bubbles to the incubation cavity. The small-volume bioreactor permits long term maintenance of tissue differentiation states in cultures. Also provided are methods of incubating cells or tissues using thebioreactor including methods of creating molecular profiles of biological effects of chemical compositions on differentiated cell or tissue samples maintained in long term culture.

Description

technical field [0001] The present invention relates to bioreactors for incubating one or more cell cultures, tissue biopsy material, cell clusters, tissue-like structures, "prototissue" or similar samples. Background technique [0002] During "classical" cell culture in essentially flat dishes, cells in general and biopsies in particular tend to dedifferentiate. Apparently, the biopsies exhibited a 'melting ice-cream effect', with cells migrating from the tissue block to the flat support surface of the petri dish. Gene expression is altered in these "migratory" cells, and they begin to behave biochemically like isolated cells rather than like cellular components of differentiated tissues. Dedifferentiated cells express biochemical pathways that differ from those normally expressed by corresponding cells in intact organisms. [0003] In contrast to "classical" cell culture conditions, "microgravity" conditions preserve a differentiated state for many types of cells in cult...

Claims

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Application Information

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IPC IPC(8): C12M3/02C12M3/06G01N33/50C12Q1/68C12N5/06C12M3/04C12N5/071C12N5/09
CPCC12M29/04C12M23/16C12M27/10
Inventor P·M·拉森S·J·费伊
Owner DRUGMODE APS
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