US7323333B2
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Materials and methods for regulating process formation in cell culture
Assignee: University of Chile
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Filing: 2004-03-31
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Granted: 2008-01-29
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Expires: 2024-03-31
Abstract
Abstract The subject invention pertains to materials and methods for inhibiting process formation and extension by cells in culture. The subject invention further includes cultures of process-forming cells wherein formation and extension of processes have been inhibited. In another aspect, the subject invention concerns methods of transplantation using process-forming cells that have been cultured by the process-inhibiting methods of the invention.
Inventors
- 👤 Pablo Caviedes
- 👤 Raul Caviedes
- 👤 Thomas B. Freeman
- 👤 Juan A. Asenjo
- 👤 Barbara A. Andrews
- 👤 Dario Sepúlveda
- 👤 Christian Arriagada
- 👤 Julio Salazar Rivera
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-22 22:55
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0618; C12N5/0619; A; A61; A61P; A61P43/00; A61K; A61K35/00; A61K35/12; C12N2533/00; C12N2533/30; C12N2533/90; C12N2533/92; Y; Y02; Y02E; Y02E50/00; Y02E50/10
Claims Analysis
Total Claims: 39
Claim 1
Independent Claim
70 words
A cell culture comprising: process-forming neurons or glial cells of the central nervous system; a cell culture medium; and a solid substrate supporting said cell culture medium; wherein said neurons or glial cells lack processes, are clustered into one or more aggregates suspended in said cell culture medium, and are not attached to said substrate; and wherein said cell culture medium has a calcium concentration of 100 uM or less.
Claim 2
Dependent Claim
70 words
A cell culture comprising: process-forming neurons or glial cells of the central nervous system; a cell culture medium; and a solid substrate supporting said cell culture medium; wherein said neurons or glial cells lack processes, are clustered into one or more aggregates suspended in said cell culture medium, and are not attached to said substrate; and wherein said cell culture medium has a calcium concentration of 100 uM or less.
Claim 3
Dependent Claim
24 words
The cell culture of claim 1 , wherein said solid substrate comprises polystyrene and has an untreated surface for supporting said cell culture medium.
Claim 4
Dependent Claim
30 words
The cell culture of claim 2 , wherein said solid substrate is a culture vessel selected from the group consisting of a Petri dish, flask, bottle, plate, tube, and vial.
Claim 5
Dependent Claim
14 words
The cell culture of claim 1 , wherein said solid substrate comprises untreated plastic.
Claim 6
Dependent Claim
26 words
The cell culture of claim 1 , wherein said solid substrate has a surface supporting said cell culture medium, and wherein said surface lacks charged molecules.
Claim 7
Dependent Claim
21 words
The cell culture of claim 1 , wherein said cell culture medium has a calcium concentration of 50 uM or less.
Claim 8
Dependent Claim
23 words
The cell culture of claim 1 , further comprising process-forming cells other than said neurons or glial cells of the central nervous system.
Claim 9
Dependent Claim
11 words
The cell culture of claim 1 , further comprising non-process-forming cells.
Claim 10
Dependent Claim
19 words
The cell culture of claim 1 , wherein said cell culture medium lacks calcium ion as a formulated component.
Claim 11
Dependent Claim
28 words
The cell culture of claim 1 , wherein each of said one or more aggregates has an average diameter in the range of 150 um to 200 um.
Claim 12
Dependent Claim
31 words
The cell culture of claim 1 , wherein said neurons or glial cells within each of said one or more aggregates includes living cells that remain viable in vivo upon implantation.
Claim 13
Dependent Claim
16 words
The cell culture of claim 1 , wherein said neurons or glial cells are fully differentiated.
Claim 14
Dependent Claim
16 words
The cell culture of claim 1 , wherein said neurons or glial cells are brain cells.
Claim 15
Dependent Claim
16 words
The cell culture of claim 1 , wherein said neurons or glial cells are human cells.
Claim 16
Dependent Claim
16 words
The cell culture of claim 1 , wherein said neurons or glial cells are primary cells.
Claim 17
Dependent Claim
19 words
The cell culture of claim 1 , wherein said neurons or glial cells are cells of a cell line.
Claim 18
Dependent Claim
13 words
The cell culture of claim 1 , wherein said neurons are dopaminergic cells.
Claim 19
Dependent Claim
16 words
The cell culture of claim 1 , wherein the process-forming neurons or glial cells are neurons.
Claim 20
Dependent Claim
17 words
The cell culture of claim 1 , wherein the process-forming neurons or glial cells are glial cells.
Claim 21
Dependent Claim
66 words
A cell culture comprising: process-forming neurons or glial cells of the central nervous system; a cell culture medium; and an untreated, polystyrene microbiological plate; wherein said neurons or glial cells lack processes, are supported by said plate, are clustered into one or more aggregates, and are not attached to said plate; and wherein said cell culture medium has a calcium concentration of 100 uM or less.
Claim 22
Dependent Claim
66 words
A cell culture comprising: process-forming neurons or glial cells of the central nervous system; a cell culture medium; and an untreated, polystyrene microbiological plate; wherein said neurons or glial cells lack processes, are supported by said plate, are clustered into one or more aggregates, and are not attached to said plate; and wherein said cell culture medium has a calcium concentration of 100 uM or less.
Claim 23
Dependent Claim
16 words
The cell culture of claim 20 , wherein said neurons or glial cells are primary cells.
Claim 24
Dependent Claim
19 words
The cell culture of claim 20 , wherein said neurons or glial cells are cells of a cell line.
Claim 25
Dependent Claim
13 words
The cell culture of claim 20 , wherein said neurons are dopaminergic cells.
Claim 26
Dependent Claim
16 words
The cell culture of claim 20 , wherein the process-forming neurons or glial cells are neurons.
Claim 27
Dependent Claim
17 words
The cell culture of claim 20 , wherein the process-forming neurons or glial cells are glial cells.
Claim 28
Dependent Claim
96 words
A method for producing the cell culture of claim 1 , comprising: placing the neurons or glial cells in the cell culture medium; and culturing the neurons or glial cells for a period of time sufficient for the neurons or glial cells to cluster into one or more aggregates, thereby producing a cell culture comprising process-forming neurons or glial cells of the central nervous system, wherein the neurons or glial cells lack cell processes, are clustered into one or more aggregates, and wherein the cell culture medium has a calcium concentration of 100 uM or less.
Claim 29
Dependent Claim
15 words
The method of claim 26 , wherein the process-forming neurons or glial cells are neurons.
Claim 30
Dependent Claim
16 words
The method of claim 26 , wherein the process-forming neurons or glial cells are glial cells.
Claim 31
Dependent Claim
96 words
A method for producing the cell culture of claim 20 , comprising: placing the neurons or glial cells in the cell culture medium; and culturing the neurons or glial cells for a period of time sufficient for the neurons or glial cells to cluster into one or more aggregates, thereby producing a cell culture comprising process-forming neurons or glial cells of the central nervous system, wherein the neurons or glial cells lack cell processes, are clustered into one or more aggregates, and wherein the cell culture medium has a calcium concentration of 100 uM or less.
Claim 32
Dependent Claim
15 words
The method of claim 29 , wherein the process-forming neurons or glial cells are neurons.
Claim 33
Dependent Claim
16 words
The method of claim 29 , wherein the process-forming neurons or glial cells are glial cells.
Claim 34
Dependent Claim
41 words
A method for preparing process-forming neurons or glial cells for transplantation, comprising: providing said cell culture of claim 1 ; removing said one or more aggregates from said culture; and combining said one or more aggregates with a pharmaceutically acceptable carrier.
Claim 35
Dependent Claim
15 words
The method of claim 32 , wherein the process-forming neurons or glial cells are neurons.
Claim 36
Dependent Claim
16 words
The method of claim 32 , wherein the process-forming neurons or glial cells are glial cells.
Claim 37
Dependent Claim
41 words
A method for preparing process-forming neurons or glial cells for transplantation, comprising: providing said cell culture of claim 20 ; removing said one or more aggregates from said culture; and combining said one or more aggregates with a pharmaceutically acceptable carrier.
Claim 38
Dependent Claim
16 words
The cell culture of claim 35 , wherein the process-forming neurons or glial cells are neurons.
Claim 39
Dependent Claim
16 words
The method of claim 35 , wherein the process-forming neurons or glial cells are glial cells.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2011-01-28 | 2011-07-30 | 2012-01-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2015-01-28 | 2015-07-30 | 2016-01-29 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2019-01-28 | 2019-07-30 | 2020-01-29 | $7700.00 | Window Closed / Lapsed |