Centrifuge for separating a sample into at least two components

• US 8,747,290 B2
• Filed: 12/08/2008
• Issued: 06/10/2014
• Est. Priority Date: 12/07/2007
• Status: Expired due to Fees

• Current Assignee: Miltenyi Biotec B.V. & Co. KG (Miltenyi GmbH)
• Original Assignee: Miltenyi Biotec GmbH
• Inventors: Miltenyi, Stefan, Schimmelpfennig, Winfried, Lantow, Holger, Biehl, Martin, Kabaha, Eiad, Schulz, Jrgen, Neuschfer, Niklas Elmar
• Primary Examiner(s): CLEVELAND, TIMOTHY C
• Application Number:
  US12/745,689
• Publication Number:
  US 20100311559A1
• Time in Patent Office: 2,010 Days
• Field of Search: 494/5, 494/10, 494/43, 494/56, 494/64, 494/67
• US Class Current: 494/10
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4615 Dendritic cells
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/4622 Antigen presenting cells
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/46434 Antigens related to induction of tolerance to non-self
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61K 39/464838 Viral antigens
• CPC Class Codes: A61M 1/0209 Multiple bag systems for separating or storing blood components
• CPC Class Codes: A61M 1/3618 Magnetic separation
• CPC Class Codes: A61M 1/3623 Means for actively controlling temperature of blood
• CPC Class Codes: A61M 1/3633 Blood component filters, e.g. leukocyte filters
• CPC Class Codes: A61M 1/3679 by absorption A61M1/3675 takes precedence
• CPC Class Codes: A61M 1/3692 Washing or rinsing blood or blood constituents
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: A61M 1/3696 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: A61M 2202/0071 product to be retained or harvested, e.g. by pheresis
• CPC Class Codes: A61M 2202/0413 Blood
• CPC Class Codes: A61M 2202/0427 Platelets; Thrombocytes
• CPC Class Codes: A61M 2202/10 Bone-marrow
• CPC Class Codes: B01D 15/3809 : of the antigen-antibody type, e.g. protein A, G, L chromatography
• CPC Class Codes: B01D 15/3885 : Using electrical or magnetic means
• CPC Class Codes: B01D 21/26 : Separation of sediment aided by centrifugal force or centripetal forcecentrifuges B04B; cyclones B04C
• CPC Class Codes: B01D 21/262 : by using a centrifuge
• CPC Class Codes: B01L 3/5021 : Test tubes specially adapted for centrifugation purposes centrifuges B04B5/04
• CPC Class Codes: B03C 1/002 : High gradient magnetic separation
• CPC Class Codes: B03C 1/027 : with reciprocating canisters
• CPC Class Codes: B03C 1/032 : Matrix cleaning systems
• CPC Class Codes: B03C 1/0332 : using permanent magnets
• CPC Class Codes: B03C 1/0335 : using coils
• CPC Class Codes: B03C 1/034 : characterised by the matrix elements
• CPC Class Codes: B03C 1/288 : disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 2201/18 : Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 : for use in medical applications
• CPC Class Codes: B04B 1/12 : with continuous discharge
• CPC Class Codes: B04B 13/00 : Control arrangements specially designed for centrifuges; Programme control of centrifuges control arrangements for feed, charge, or discharge B04B11/00
• CPC Class Codes: B04B 2005/0464 : with hollow or massive core in centrifuge bowl
• CPC Class Codes: B04B 2013/006 : Interface detection or monitoring of separated components
• CPC Class Codes: B04B 5/00 : Other centrifuges
• CPC Class Codes: B04B 5/04 : Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
• CPC Class Codes: B04B 5/0442 : with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: B04B 5/10 : Centrifuges combined with other apparatus, e.g. electrostatic separators; Sets or systems of several centrifuges B04B5/12 takes precedence; magnetic or electrostatic separators B03C; amassing particles by electric fields, e.g. by agglomeration B03C3/0175
• CPC Class Codes: B04B 7/08 : Rotary bowls centrifugal casting machines B22D
• CPC Class Codes: C12M 23/28 : disposable or single use
• CPC Class Codes: C12M 33/10 : by centrifugation centrifuges in general B04B; Cyclones cyclones in general B04C
• CPC Class Codes: C12M 41/48 : Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12M 47/04 : Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 2500/34 : Sugars
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 5/0018 : Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12N 5/0646 : Natural killers cells [NK], NKT cells
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0669 : Bone marrow stromal cells; Whole bone marrow isolated stem cells from bone marrow C12N5/0647, C12N5/0663
• CPC Class Codes: G01N 2333/70596 : Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 33/58 : involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: G01N 35/0098 : involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C

Abstract

The invention relates to a centrifuge for separating a sample into at least two components, comprising a chamber for receiving a sample to be centrifuged. According to the invention, the centrifuge further comprises a means for controlling the progress of the sample separation is located at the chamber.

Claims

1. A centrifuge chamber configured for separating a sample of cells into at least two portions, wherein the chamber comprises:

(a) a circular base plate, the center of which is substantially perpendicular to a rotational axis of the chamber,

(b) a cladding which is substantially perpendicular to the base plate,

(c) a circular cover plate which is positionable on an edge of the cladding that is remote from the base plate,

(d) an interior configured for processing the sample of cells,

(e) a chamber window in the base plate or the cover plate through which a part of the sample may be monitored during centrifugation, wherein the chamber window extends radially in the base plate or cover plate from a position near the rotation axis to a position near the perimeter; and

(f) a detection channel projecting into the base plate or the cover plate from the interior of the chamber to the chamber window, wherein the detection channel is positioned radially from the interior of the chamber to and along the window, and is configured to receive and show the part of the sample through the window during centrifugation;

wherein the chamber is configured for use in a centrifuge that comprises a detector, wherein the detector is configured and aligned to monitor separation of cells in the part of the sample showing through and along the chamber window.

2. The centrifuge chamber according to claim 1,

wherein the chamber window is reflective so as to receive light from a light source, direct the light through the part of the sample that appears in the detection channel, and then redirect light received from the part of the sample to the detector.

3. The centrifuge chamber according to claim 2, wherein

the chamber window comprises a prism oriented radially over the detection channel from a position near the rotation axis to a position near the perimeter.

4. The centrifuge chamber according to claim 1, wherein the interior is configured such that the centrifuge chamber can serve as a container for cultivation of cells during centrifugation.

5. The centrifuge chamber according to claim 1, which further comprises at least one layered structure configured for adhering and growing cells thereon.

6. The centrifuge chamber according to claim 1, which is disposable.

7. The centrifuge chamber according to claim 1, which comprises a material chosen from plastics, polystyrol (PS), polystyrene, polyvinylchloride, polycarbonate, glass, polyacrylate, polyacrylamide, polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polytetrafluorethylene (PTFE), thermoplastic polyurethane (TPU), silicone, polyethylene (PE), collagen, chitin, alginate, hyaluronic acid derivatives, polylactide (PLA), polyglycolide (PGA) and their copolymers, polystyrol (PS), polysterene, polycarbonate, polyacrylate, ceramics, hydroxyapatite, calcium phosphate, and compositions comprising one or more of such materials.

8. A method for separating a sample into at least two components, comprising providing a sample that can be separated into at least two components, and centrifuging the sample in a centrifuge chamber according to claim 1.

9. The method according to claim 8, further comprising:

detecting the progress of separation by detecting the formation of layers of the sample, the change of pH value, and/or the change of temperature.

10. The method according to claim 8, whereby the sample comprises blood cells or bone marrow cells.

11. The centrifuge chamber according to claim 1, wherein the base plate comprises one or more openings for delivery of gases into the chamber during centrifugation.

12. The centrifuge chamber according to claim 1, wherein one or more sensor pads are located on the base plate.

13. The centrifuge chamber according to claim 1, further comprising a sample inlet port and an opening for air or gas,

wherein the centrifuge chamber is configured such that the sample can pass through the chamber by way of the sample inlet port, and air or gas to support culturing of cells in the chamber can pass through the chamber by way of the opening, both without stopping centrifugation of the chamber.

14. The centrifuge chamber according to claim 1, comprising a plurality of layered structures that are separate from the base and cover plates and are perpendicular to the rotational axis upon which cells may be cultured.

15. The centrifuge chamber according to claim 14, which is configured such that cells adhering to the layered structures may be separated from cells with lesser affinity for the layered structures.

16. The centrifuge chamber according to claim 14, wherein the layered structures are configured for immunoaffinity separation of cells in the chamber.

17. The centrifuge chamber according to claim 14, wherein the layered structures are configured for magnetic separation of cells in the chamber.

18. The centrifuge chamber according to claim 13, having a system of channels for gas flow assembled in the chamber in a spiral pattern.

19. The centrifuge chamber according to claim 11, further comprising a hydrophobic membrane that is configured to separate gas that has entered the chamber through the one or more openings from sample being processed in the chamber.

20. The centrifuge chamber according to claim 1, further comprising a central line passing into the chamber through and along the rotational axis, and connected to at least one compartment of the chamber, wherein the chamber is configured so that the sample being processed can pass from outside the chamber into or out of the compartment through the central line during centrifugation.

21. The centrifuge chamber according to claim 20, further comprising an outer line that is parallel to and concentric with the central line through and along the rotational axis.

22. The centrifuge chamber according to claim 1, which can be sterilized.

23. A centrifuge comprising:

(1) a centrifuge chamber according to claims 1; and

(2) a detector configured and aligned to monitor separation of cells in the part of the sample showing through and along the chamber window.

24. The centrifuge according to claim 23, further comprising a light source configured and aligned to direct an illuminating beam onto the part of the sample that appears in the detection channel through the chamber window during centrifugation, and thereafter to the detector, wherein the detector comprises an optical sensor.

25. The centrifuge according to claim 24, wherein the illuminating beam from the light source is essentially perpendicular to the rotational axis of the chamber.