US4734372A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Cell culturing methods and apparatus
Assignee: NU-TECH BIO-MED Inc
•
Filing: 1984-06-21
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Granted: 1988-03-29
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Expires: 2004-06-21
Abstract
Abstract A system and protocol for performing cytotoxicity studies including apparatus and methods for culturing the biopsied cells, for providing oxygenated nutrients, for removing cell-waste products, for introducing a fluorogenic substrate, for introducing cytotoxic agents including anticancer drugs, for measuring the released fluorescence and for measuring intracellular accumulation of fluorescein. Cytotoxicity can be determined by measuring fluorescence in the efflux of the vessel and concommitantly by direct photometric comparisons of the cells in the vessel before and after exposure to the cytotoxic agent.
Inventors
- 👤 M. Boris Rotman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 12:38
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/5011; G01N33/5014; G01N2800/00; G01N2800/52
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
126 words
A cytotoxicity assay system comprising a cell culturing vessel having a cell enclosure into which cells can be introduced and cultured; nutrient-transporting means for transporting nutrients to the cells, the nutrient-transporting means include a source of nutrients and at least one fluid permeable membrane, connecting said source of nutrients to said cells, across which nutrients can migrate to nourish the cells within the vessel; fluorogenic substrate-transporting means for transporting a fluorogenic substrate to the cell compartment, so that living cells within the vessel will take up said substrate and accumulate a characteristic fluorescence; means for introducing a therapeutic agent into the cell compartment; and means for measuring cytotoxicity within the cell compartment, including means for detected changes in the fluorescence of the cells within the vessel.
Claim 2
Dependent Claim
126 words
A cytotoxicity assay system comprising a cell culturing vessel having a cell enclosure into which cells can be introduced and cultured; nutrient-transporting means for transporting nutrients to the cells, the nutrient-transporting means include a source of nutrients and at least one fluid permeable membrane, connecting said source of nutrients to said cells, across which nutrients can migrate to nourish the cells within the vessel; fluorogenic substrate-transporting means for transporting a fluorogenic substrate to the cell compartment, so that living cells within the vessel will take up said substrate and accumulate a characteristic fluorescence; means for introducing a therapeutic agent into the cell compartment; and means for measuring cytotoxicity within the cell compartment, including means for detected changes in the fluorescence of the cells within the vessel.
Claim 3
Dependent Claim
25 words
The system of claim 1 wherein the vessel further includes at least one transparent surface through which fluorescence within the cell enclosure can be observed.
Claim 4
Dependent Claim
30 words
The system of claim 1 wherein the vessel further includes at least one sampling port for withdrawing an sample from the cell enclosure to measure the fluorescence of the sample.
Claim 5
Dependent Claim
22 words
The system of claim 1 wherein the fluid permeable membrane has a pore size ranging from about 5 to about 35 microns.
Claim 6
Dependent Claim
14 words
The system of claim 1 wherein the fluid permeable membrane is a synthetic capillary.
Claim 7
Dependent Claim
15 words
The system of claim 1 wherein the fluid permeable membrane is a fibrous cellulose material.
Claim 8
Dependent Claim
12 words
The system of claim 1 wherein the permeable membrane is polymeric mesh.
Claim 9
Dependent Claim
14 words
The system of claim 1 wherein the means for measuring fluorescence is a camera.
Claim 10
Dependent Claim
21 words
The system of claim 8 wherein the means for measuring fluorescence further includes a monochromatic light source and a matched filter.
Claim 11
Independent Claim
75 words
A method of assaying the sensitivity of biopsied cells to therapeutic agents, the method comprising: a. culturing the cells in a vessel; b. contacting the cells with a fluorogenic substrate whereby living cells accumulate a characteristic amount of fluorescence; c. introducing the agent into the vessel; and d. measuring changes in the fluorescence by photometrically analyzing the intensity of fluorescence within the vessel as an indicator of the sensitivity of the cells to the agent.
Claim 12
Independent Claim
75 words
A method of assaying the sensitivity of biopsied cells to therapeutic agents, the method comprising: a. culturing the cells in a vessel; b. contacting the cells with a fluorogenic substrate whereby living cells accumulate a characteristic amount of fluorescence; c. introducing the agent into the vessel; and d. measuring changes in the fluorescence by photometrically analyzing the intensity of fluorescence within the vessel as an indicator of the sensitivity of the cells to the agent.
Claim 13
Dependent Claim
23 words
The method of claim 10 wherein the step of measuring changes in fluorescence further including photometrically recording the fluorescence with a photographic camera.
Claim 14
Dependent Claim
23 words
The method of claim 10 wherein the step of measuring changes in fluorescence further includes photometrically recording the fluorescence with a video camera.
Claim 15
Dependent Claim
23 words
The method of claim 10 wherein the step of measuring changes in fluorescence further includes illuminating the cultured cells with blue monochromatic light.
Claim 16
Dependent Claim
26 words
The method of claim 13 wherein the step of measuring changes in fluorescence further includes filtering light from the cultured cells with a matching blue filter.
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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) | 1991-03-29 | 1991-09-28 | 1992-03-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-03-29 | 1995-09-28 | 1996-03-29 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-03-29 | 1999-09-28 | 2000-03-29 | $7700.00 | Window Closed / Lapsed |