US4559299A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Cytotoxicity assays in cell culturing devices
Assignee: BROWN UNIVERSITY RESEARCH FOUNDATION 42 CHARLESFIELD STREET PROVIDENCE RHODE ISLAND 02912 A RHODE ISLAND CORP
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Filing: 1983-02-04
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Granted: 1985-12-17
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Expires: 2003-02-04
Abstract
Abstract Methods and devices for assaying the sensitivity of of biopsied cells to therapeutic agents are disclosed. Cells are cultured in artificial organs and then contacted with a fluorogenic substrate such that living cells accumulate a characteristic amount of fluorescence. The agent is then introduced into the organ and changes in the fluorescence released by the cells serve as an indicator of the sensitivity of the cells to the agent.
Inventors
- 👤 M. Boris Rotman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:52
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/5011; C; C12; C12M; C12M23/00; C12M23/02; C12M23/16; C12M29/00; C12M29/04; C12M29/10; C12M41/00; C12M41/46; Y; Y10; Y10S; Y10S435/00; Y10S435/808; Y10S435/81; Y10S435/811; Y10T; Y10T436/00; Y10T436/25; Y10T436/25375; Y10T436/255
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
108 words
A method of assaying the sensitivity of biopsied cells to therapeutic agents, the method comprising: (a) culturing the cells in a cell culturing apparatus which comprises either at least one semi-permeable membrane surface or at least one semi-permeable capillary vessel which enables the perfusion of nutrients to the biopsied cells; (b) contacting the cells with a fluorogenic substrate, whereby living cells release an initial characteristic amount of fluorescence; (c) measuring said initial fluorescence to establish a baseline; (d) introducing the agent into the culturing apparatus; and (e) measuring changes in the fluorescence released by the cells as an indicator of the sensitivity of the cells to the agent.
Claim 2
Independent Claim
108 words
A method of assaying the sensitivity of biopsied cells to therapeutic agents, the method comprising: (a) culturing the cells in a cell culturing apparatus which comprises either at least one semi-permeable membrane surface or at least one semi-permeable capillary vessel which enables the perfusion of nutrients to the biopsied cells; (b) contacting the cells with a fluorogenic substrate, whereby living cells release an initial characteristic amount of fluorescence; (c) measuring said initial fluorescence to establish a baseline; (d) introducing the agent into the culturing apparatus; and (e) measuring changes in the fluorescence released by the cells as an indicator of the sensitivity of the cells to the agent.
Claim 3
Dependent Claim
38 words
The method of claim 1 wherein the step of measuring changes in fluorescence further comprises comparing the baseline measurement with a second measurement of released fluorescence after a second contact with the substrate subsequent to introducing the agent.
Claim 4
Dependent Claim
17 words
The method of claim 1 wherein changes in fluorescence are determined by measuring changes in fluorescent activity.
Claim 5
Dependent Claim
12 words
The method of claim 1 wherein changes in fluorescence are measured kinetically.
Claim 6
Dependent Claim
23 words
The method of claim 1 wherein the method further comprises varying the temperature of the culturing apparatus prior to measuring changes in fluorescence.
Claim 7
Dependent Claim
21 words
The method of claim 1 wherein the fluorogenic substrate is a fluorescein containing compound which liberates fluorescein when hydrolyzed by cells.
Claim 8
Dependent Claim
34 words
The method of claim 1 wherein cells are contacted with at least one substrate chosen from the group of fluorescein diacetate, fluorescein monoacetate and derivatives thereof, wherein fluorescein is liberated when hydrolyzed by cells.
Claim 9
Dependent Claim
27 words
The method of claim 1 wherein the cell culturing apparatus comprises at least one semi-permeable capillary vessel which enables the perfusion of nutrients to the biopsied cells.
Claim 10
Dependent Claim
27 words
The method of claim 1 wherein the cell culturing apparatus comprises at least one semi-permeable membrane surface which enables the perfusion of nutrients to the biopsied cells.
Claim 11
Dependent Claim
39 words
The method of claim 1 wherein the artificial cell culturing apparatus comprises a cell compartment which provides for influx of an oxygenated medium and changes in the fluorescence are detected by changes in nutrient medium exiting the cell compartment.
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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) | 1988-12-16 | 1989-06-17 | 1989-12-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-12-16 | 1993-06-17 | 1993-12-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-12-16 | 1997-06-17 | 1997-12-17 | $7700.00 | Window Closed / Lapsed |