Microfabricated apparatus for cell based assays

• US 7,935,522 B2
• Filed: 04/04/2006
• Issued: 05/03/2011
• Est. Priority Date: 04/27/1998
• Status: Expired due to Term

• Current Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Original Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Inventors: Thomas, Nicholas
• Primary Examiner(s): Beisner; William H
• Application Number:
  US11/397,401
• Publication Number:
  US 20060194273A1
• Time in Patent Office: 1,855 Days
• Field of Search: 435/288.5, 435/293.1, 435/297.5, 422/415, 422/72, 422/506
• US Class Current: 435/288.5
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0806 Standardised forms, e.g. compact disc [CD] format
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 23/38 Caps; Covers; Plugs; Pouring means
• CPC Class Codes: C12M 25/16 Particles; Beads; Granular material; Encapsulation chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00

Abstract

Apparatus and methods are provided for performing cell growth and cell based assays in a liquid medium. The apparatus comprises a base plate supporting a plurality of micro-channel elements, each micro-channel element comprising a cell growth chamber, an inlet channel and an outlet channel, a cover plate positioned over the base plate to define the chambers and connecting channels. Means are incorporated in the cell growth chambers, for cell attachment and cell growth. In particular, the invention provides a rotatable disc microfabricated for performing cell growth and cell based assays. The apparatus and method can be used for the growth of cells and the detection and measurement of a variety of biochemical processes and products using non-invasive techniques, that is techniques which do not compromise the integrity or viability of cells.

Claims

1. Apparatus microfabricated for performing cell growth and cell based assays in a liquid medium, said apparatus comprising:

a) a base plate supporting a plurality of micro-channel elements, each comprising a cell growth chamber, an inlet channel for supplying liquid sample to said chamber and an outlet channel for removal of liquid sample from said chamber;

b) a cover plate positioned over said base plate said cover plate extending over said elements so as to define said chambers and connecting channels; and

c) a plurality of holes in said apparatus in communication with said channels for supplying liquid sample to said chamber and removal of liquid sample from said chamber,

wherein one or more components of the apparatus comprise a gas permeable film or membrane adapted to allow the use of CO₂-buffered media in the cell growth chamber and to allow oxygen to permeate to the cells for their metabolism during growth, wherein at least some of said micro-channel elements each further comprises one or more assay chambers for performing assays involving cellular constituents, the assay chambers being connected in line between said cell growth chamber and said outlet channel.

2. Apparatus according to claim 1 wherein said base plate comprises a rotatable disc comprising a sample introduction port located towards the centre of the disc, the sample introduction port being connected to an annular sample reservoir, and wherein said micro-channel elements are radially dispersed on said disc with their respective input channels connected to said reservoir and adapted to receive a sample from said reservoir.

3. Apparatus according to claim 1, wherein said cover plate is comprises a gas permeable plastic material.

4. Apparatus according to claim 1, further comprising at least a portion of a surface of said cell growth chamber that is adapted to allow for cell attachment to the surface portion.

5. Apparatus according to claim 1, wherein the cell growth chamber further comprises one or more microcarrier beads located in said chamber, wherein each of said microcarrier beads is capable of supporting cell attachment.

6. Apparatus according to claim 1, wherein the chamber further includes raised moulded features disposed on the base portion of the cell growth chamber to form pillars.

7. Apparatus according to claim 1, wherein the cross-sectional area of said inlet channel is greater than that of said outlet channel.

8. Apparatus according to claim 1, wherein the cross-sectional area of said outlet channel is between 0.99 and 0.01 times that of said inlet channel.

9. Apparatus according to claim 1, wherein the assay chamber or chambers are connected to each other and to said cell growth chamber by an intermediate channel or channels in the order of:

inlet channel, cell growth chamber, intermediate channel, assay chamber or chambers, outlet channel, and wherein the cross-sectional areas of the respective channels reduce progressively from the inlet channel to the outlet channel.

10. Apparatus according to claim 9, wherein the cross-sectional area of the or each intermediate channel and the outlet channel is between 0.99 and 0.01 times that of the immediately preceding channel.

11. Apparatus according to claim 9, wherein there is provided in or on an interior surface of one or more of said assay chambers a layer comprising a scintillant substance.

12. Apparatus according to claim 11, wherein the layer comprising a scintillant substance includes a binding moiety bound thereto, said binding moiety being a member of a specific binding pair selected from the group consisting of biotin, streptavidin, protein A, antibodies, lectins, hormone-receptors, nucleic acid probes, and DNA-binding proteins.

13. Apparatus according to claim 1, further comprising a suspension of cells to be grown in each of said chambers.

14. Apparatus according to claim 13, wherein said cells are adherent to said cell growth chambers when cultured therein.

15. Apparatus according to claim 1, wherein said cells derive from a species selected from the group consisting of human, rodents and simians.

16. Apparatus according to claim 1, wherein said cells are eukaryotic.

17. The apparatus of claim 1, further comprising an evaporation prevention component sealing the plurality of openings in the apparatus, the component being adapted to prevent evaporation of a liquid within the apparatus without interfering with gas permeation through the apparatus.

18. An apparatus comprising:

a) a first element affixed to a second element, wherein one or both of the elements comprise a molded structure that defines in the apparatus a plurality of micro-channel elements, each micro-channel element comprising;

i) a cell growth chamber having at least a portion of a surface of the cell growth chamber that is adapted to allow for cell attachment to the surface portion,

ii) a first channel connected to the cell growth chamber, the first channel being capable of use for the delivery of liquid into the cell growth chamber,

iii) and a second channel connected to the cell growth chamber, the second channel being capable of use for the removal of liquid from the cell growth chamber,

iv) a plurality of openings in the apparatus communicating with the first or second channels,

wherein one or more components of the apparatus comprise a gas permeable film or membrane adapted to allow the use of CO₂-buffered media in the cell growth chamber and to allow oxygen to permeate to the cell for its metabolism during growth, and wherein at least some of said micro-channel elements each further comprises one or more assay chambers for performing assays involving cellular constituents, the assay chambers being connected in line between said cell growth chamber and an outlet channel.

19. The apparatus of claim 18, further comprising an evaporation prevention component sealing the plurality of openings in the apparatus, the component being adapted to prevent evaporation of a liquid within the apparatus without interfering with gas permeation through the apparatus.