Device for preparing an undifferentiated cell from a more committed cell

• US 8,163,536 B2
• Filed: 05/09/2001
• Issued: 04/24/2012
• Est. Priority Date: 05/10/2000
• Status: Expired due to Fees

• Current Assignee: Tristem Trading (Cyprus) Limited
• Original Assignee: Tristem Trading (Cyprus) Limited
• Inventors: Abuljadayel, Ilham Mohamed Saleh Saeed
• Primary Examiner(s): CANELLA, KAREN A
• Application Number:
  US09/853,188
• Publication Number:
  US 20030064503A1
• Time in Patent Office: 4,003 Days
• Field of Search: 435/286.1, 435/286.4, 435/286.5, 435/286.6, 435/287.3, 435/303.1
• US Class Current: 435/284.1
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C12M 23/04 Flat or tray type, drawers C12M23/10, C12M23/12, C12M23/16 take precedence
• CPC Class Codes: C12M 23/26 flexible flexible containers for laboratory use B01L3/505
• CPC Class Codes: C12M 27/02 Stirrer or mobile mixing elements
• CPC Class Codes: C12M 41/14 Incubators; Climatic chambers per se B01L1/00
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2500/24 Iron; Fe chelators; Transferrin
• CPC Class Codes: C12N 2501/14 Erythropoietin [EPO]
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/50 Cell markers; Cell surface determinants
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

A device for preparing an undifferentiated cell, the device comprises means for contacting a more committed cell with an agent that causes the more committed cell to retrodifferentiate into an undifferentiated cell.

Claims

1. A device for forming and/or increasing the relative number of undifferentiated cells in a cell population comprising haematopoietic cells, including committed cells, which device comprises:

(i) a chamber;

(ii) an input storage container containing a cell population including committed cells;

(iii) means for introducing into the chamber cells of the cell population including committed cells;

(iv) transfer means between the input storage container and the chamber for transferring the cells from the storage container continuing to the chamber;

(v) means for introducing into the chamber an agent selected from the group consisting of;

(a) an antibody that binds to MHC antigens,

(b) erythropoietin, and

(c) GM-CSF;

(vi) a further transfer means for transferring a volume of the agent into the chamber, including a reservoir of the agent;

(vii) incubation means for incubating said committed cells in the presence of the agent;

(viii) mixing means for mixing the agent and the cell population in the chamber;

(ix) an output storage container; and

(x) means for removing from the chamber into the output storage container a sample of cells comprising undifferentiated cells;

wherein the chamber, the input storage container, the further transfer means, and the output storage container are disposable.

3. A device according to claim 1 wherein said device comprises:

measuring means for measuring the volume of said cell population; and

/or

means for conducting cell counts and for measuring the cell concentration of said cell population; and

/or

calculator means for calculating the volume of agent to be added to the chamber; and

/or

carbon dioxide control means for controlling the concentration of carbon dioxide in said chamber; and

/or

temperature control means for controlling the temperature in said chamber; and

/or

timing means for timing the incubation period; and

/or

display means for displaying to the user the remaining time period of the incubation period; and

/or

alarm means for alerting the user of completion of the incubation period; and

/or

harvesting means for harvesting cells from the chamber by dislodging cells that are attached to the surface of the chamber; and

/or

removal means for removing a sample of cells, comprising undifferentiated cells, from the chamber into a storage container; and

/or

sealing means for sealing a storage container comprising a population of cells comprising undifferentiated cells; and

/or

communicating means for the device to remotely communicate orders and/or confirm that operations are being or have been performed correctly,

wherein the transfer means is configurable to transfer a pre-determined amount of the cell population from the storage container to the chamber; and

wherein the means for introducing an agent into the chamber comprises the further transfer means for transferring a volume of agent to the chamber, and/or for transferring a calculated volume of agent to the chamber.

4. A device according to claim 3 wherein the means for conducting cell counts is a coulter counter.

5. A device according to claim 3 wherein the means for conducting cell counts is a cytometer.

6. A device according to claim 3 wherein the further transfer means is configurable to transfer a volume of agent to the chamber and comprises a syringe driven by a motor.

7. A device according to claim 3 wherein the further transfer means is configurable to transfer a calculated volume of agent to the chamber and comprises a syringe driven by a motor.

8. A device according to claim 3 wherein the harvesting means harvests the undifferentiated cells from the chamber.

9. A device according to claim 3 wherein the communicating means includes a microprocessor to collect and/or store data pertaining to agent(s) increasing the relative number of undifferentiated cells in a cell population, and/or ordering a supply thereof and/or operations and modem means for transmitting such data.

10. A device according to any one of claims 1-3 wherein the committed cells are non-cancer cells.

11. A device according to any one of claims 1-3 wherein the committed cells are differentiated cells.

12. A device according to any one of claims 1-3 wherein the committed cells are selected from CFC-T cells, CFC-B cells, CFC-Eosin cells, CFC-Bas cells, CFC-GM cells, CFC-MEG cells, CFC-E cells, T cells and B cells.

13. A device according to any one of claims 1-3 wherein the undifferentiated cells are pluripotent stem cells.

14. A device according to any one of claims 1-3 wherein the undifferentiated cells are characterised by one or more of the following cell surface marker designations:

CD34+, HLA-DR−

, CD38−

, CD117, AC133, CD90 and/or CD45low.

15. A device according to any one of claims 1-3 wherein the undifferentiated cells are MHC class I+ and/or MHC class II+ cells.

26. A device according to any one of claims 1-3 wherein the cell population including committed cells is a buffy coat blood sample or is from a buffy coat blood sample.

2. A device for forming and/or increasing the relative number of undifferentiated cells in a cell population comprising haematopoietic cells, including committed cells, which device comprises:

a chamber;

(ii) an input storage container containing a cell population including committed cells;

(iii) means for introducing into the chamber cells of the cell population including committed cells;

(iv) transfer means between the input storage container and the chamber for transferring the cells from the storage container continuing to the chamber;

(v) means for introducing into the chamber an agent selected from the group consisting of;

(a) an antibody that binds to MHC antigens,

(b) erythropoietin, and

(c) GM-CSF;

(vi) a further transfer means for transferring a volume of the agent to the chamber, wherein the further transfer means comprises a volume of the agent;

(vii) incubation means for incubating the agent and said committed cells;

(viii) mixing means for mixing the agent and the cell population in the chamber; and

(ix) means for removing from the chamber into an output storage container a sample of cells comprising undifferentiated cells;

wherein the chamber, the input storage container, the further transfer means, and the output storage container are disposable.

16. A device according to claim 2 wherein the antigen is an MHC class I antigen or an MHC class II antigen.

17. A device according to claim 16 wherein the class I antigen is an HLA-A receptor, an HLA-B receptor, an HLA-C receptor, an HLA-E receptor, an HLA-F receptor or an HLA-G receptor and said class II antigen is an HLA-DM receptor, an HLA-DP receptor, an HLA-DQ receptor or and HLA-DR receptor.

18. A device according to claim 17 wherein the antigen is an HLA-DR receptor.

19. A device according to claim 2 wherein the antigen comprises a β

-chain having homologous regions.

20. A device according to claim 19 wherein the antigen comprises at least the homologous regions of the β

-chain of HLA-DR.

21. A device according to claim 2 wherein the antibody is a monoclonal antibody to the MHC antigen.

22. A device according to claim 2 wherein the antibody is selected from the group consisting of monoclonal antibody CR3/43 and the monoclonal antibody TAL 1B5.

23. A device according to claim 21 wherein the antibody is selected from the group consisting of monoclonal antibody CR3/43 and the monoclonal antibody TAL 1B5.

24. A device according to claim 2 wherein the agent modulates MHC gene expression.

25. A device according to claim 24 wherein the agent modulates MHC class I+ and/or MHC class II+ expression.

27. A device for forming and/or increasing the relative number of undifferentiated cells in a cell population comprising haematopoietic cells, which device comprises:

(i) a chamber;

(ii) an input storage container containing a cell population including committed cells;

(iii) means for introducing into the chamber cells of the cell population including haematopoietic cells;

(iv) transfer means between the input storage container and the chamber for transferring the cells from the storage container continuing to the chamber;

(v) means for introducing an agent into the chamber, wherein the agent is selected from the group consisting of;

(a) an antibody that binds to MHC antigens, (b) erythropoietin, and (c) GM-CSF;

(vi) a further transfer means for transferring a volume of the agent to the chamber, wherein the further transfer means comprises a volume of the agent;

(vii) incubation means for incubating said committed cells in the presence of the agent;

(viii) mixing means for mixing the agent and the cell population in the chamber;

(ix) an output storage container; and

(x) means for removing from the chamber into the output storage container a sample of cells comprising undifferentiated cells;

wherein the chamber, the input storage container, the further transfer means, and the output storage container are disposable.

28. A device for forming and/or increasing the relative number of cells having a cell surface marker designation CD34+ and/or HLA-DR−

and/or CD38− and

/or CD117 and/or AC133 and/or CD90 and/or CD45low in a cell population comprising haematopoietic cells, including committed cells, which device comprises;

(i) a chamber;

(ii) an input storage container containing a cell population including committed cells;

(iii) means for introducing into the chamber cells of the cell population including committed cells;

(iv) transfer means between the input storage container and the chamber for transferring the cells from the storage container continuing to the chamber;

(v) means for introducing an agent into the chamber;

(vi) a further transfer means for transferring a volume of the agent to the chamber, wherein the further transfer means comprises a volume of the agent;

(vii) incubation means operable to incubate said committed cells and said agent selected from the group consisting of an antibody that binds to MHC antigens, erythropoietin, and GM-CSF;

(viii) mixing means for mixing the agent and the cell population in the chamber;

(ix) an output storage container; and

(x) means for removing from the chamber into the output storage container a sample of cells comprising undifferentiated cells;

wherein the chamber, the input storage container, the further transfer means, and the output storage container are disposable.

29. A device for treating a starting hematopoietic cell population comprising haematopoietic cells with an agent selected from the group consisting of (a) an antibody that binds to MHC antigens, (b) erythropoietin, and (c) GM-CSF, and capable of increasing the relative number of undifferentiated cells, which device comprises:

(a) a support hook for attachment of an inlet storage container containing a starting cell population, wherein said inlet storage container is a blood bag;

(b) a heated incubation chamber;

(c) means for drawing the starting cell population from the inlet storage container into the heated incubation chamber;

(d) a refrigerated chamber for insertion of a syringe containing the agent selected from the group consisting of (a) an antibody that binds to MHC antigens, (b) erythropoietin, and (c) GM-CSF;

(e) means for delivering the agent from the refrigerated chamber into the heated incubation chamber;

(f) rotating mixer that mixes the agent and the cell population in the heated incubation chamber;

(g) a carbon dioxide control means for controlling the concentration of carbon dioxide in the heated incubation chamber;

(h) a means for hanging an outlet storage container, wherein said outlet storage container is a blood bag; and

(i) means for drawing the treated cell population from the heated incubation chamber into the outlet storage container.

30. A device according to claim 29 wherein the means for drawing the starting cell population from the inlet storage container into the heated incubation chamber comprises a peristaltic pump.

31. A device according to claim 29 wherein the means for drawing the treated cell population from the incubation chamber into the outlet storage container comprises a peristaltic pump.

32. A device according to claim 29 wherein the support hook forms part of an electronic balance for weighing the inlet storage container.

33. The device according to claim 29 wherein the means for delivering the agent from the refrigerated chamber into the heated incubation chamber comprises a stepper motor for discharging a syringe.