US5744363A
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US
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Method for establishing a tumor-cell line by preparing single-cell suspension of tumor cells from tumor biopsies
Assignee: National Institute of Immunology
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Filing: 1996-12-13
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Granted: 1998-04-28
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Expires: 2016-12-13
Abstract
Abstract The present invention provides a novel method for the establishment of tumorigenic cell lines from biopsies of human or animal tumors. The method includes preparing a single-cell suspension of a tumor cells by injecting cell culture medium into an isolated tumor or a portion thereof to flush out the tumor cells. The tumor or portion thereof may be in a cell culture maintenance medium. The step of injecting cell culture medium into the isolated tumor or portion thereof involves piercing the tumor or portion thereof with a needle to inject the cell culture medium and flush out the tumor cells. The steps of piercing the tumor or portion thereof with a needle and injecting the cell culture medium to flush out the tumor cells preferably are repeated until the single-cell suspension has a particular cell density. The method also includes establishing a primary tumor cell culture by growing the cells from the single-cell suspension in a cell culture medium.
Inventors
- 👤 Rama Mukherjee
- 👤 Manu Jaggi
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 06:10
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0693; A; A61; A61K; A61K38/00; A61K38/04; A61K38/10
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
31 words
A method of preparing a single-cell suspension of tumor cells, the method comprising injecting cell culture medium into an isolated tumor or a portion thereof to flush out the tumor cells.
Claim 2
Independent Claim
31 words
A method of preparing a single-cell suspension of tumor cells, the method comprising injecting cell culture medium into an isolated tumor or a portion thereof to flush out the tumor cells.
Claim 3
Dependent Claim
44 words
A method as claimed in claim 1, wherein the step of injecting cell culture medium into the isolated tumor or portion thereof comprises piercing the tumor or portion thereof with a needle to inject the cell culture medium and flush out the tumor cells.
Claim 4
Dependent Claim
19 words
A method as claimed in claim 1, wherein the tumor or portion thereof is in a cell culture medium.
Claim 5
Dependent Claim
19 words
A method as claimed in claim 2, wherein the tumor or portion thereof is in a cell culture medium.
Claim 6
Dependent Claim
54 words
A method as claimed in claim 2, wherein the steps of piercing the tumor or portion thereof with a needle and injecting the cell culture medium to flush out the tumor cells are repeated until the single-cell suspension has a cell density of about 10 3 cells/ml to about 10 6 cells/ml of suspension.
Claim 7
Dependent Claim
49 words
A method as claimed in claim 4, wherein the steps of piercing the tumor or portion thereof with a needle and injecting the cell culture medium to flush out the tumor cells are repeated until the single-cell suspension has a cell density of approximately 10 3 cells/ml of suspension.
Claim 8
Dependent Claim
42 words
A method of establishing a primary tumor cell culture, the method comprising the steps of: (a) preparing a single-cell suspension according to the method claimed in claim 1; and (b) growing the cells from the single-cell suspension in a cell culture medium.
Claim 9
Dependent Claim
30 words
A method of establishing a primary tumor cell culture as claimed in claim 7, wherein growing the cells in step (b) further comprises passaging the cells at least four times.
Claim 10
Dependent Claim
42 words
A method of establishing a primary tumor cell culture, the method comprising the steps of: (a) preparing a single-cell suspension according to the method claimed in claim 2; and (b) growing the cells from the single-cell suspension in a cell culture medium.
Claim 11
Dependent Claim
26 words
A method of establishing a primary tumor cell culture as claimed in claim 9, wherein growing the cells in step (b) further comprises passaging the cells.
Claim 12
Dependent Claim
42 words
A method of establishing a primary tumor cell culture, the method comprising the steps of: (a) preparing a single-cell suspension according to the method claimed in claim 3; and (b) growing the cells from the single-cell suspension in a cell culture medium.
Claim 13
Dependent Claim
26 words
A method of establishing a primary tumor cell culture as claimed in claim 11, wherein growing the cells in step (b) further comprises passaging the cells.
Claim 14
Dependent Claim
42 words
A method of establishing a primary tumor cell culture, the method comprising the steps of: (a) preparing a single-cell suspension according to the method claimed in claim 4; and (b) growing the cells from the single-cell suspension in a cell culture medium.
Claim 15
Dependent Claim
26 words
A method of establishing a primary tumor cell culture as claimed in claim 13, wherein growing the cells in step (b) further comprises passaging the cells.
Claim 16
Dependent Claim
42 words
A method of establishing a primary tumor cell culture, the method comprising the steps of: (a) preparing a single-cell suspension according to the method claimed in claim 5; and (b) growing the cells from the single-cell suspension in a cell culture medium.
Claim 17
Dependent Claim
26 words
A method of establishing a primary tumor cell culture as claimed in claim 15, wherein growing the cells in step (b) further comprises passaging the cells.
Claim 18
Dependent Claim
42 words
A method of establishing a primary tumor cell culture, the method comprising the steps of: (a) preparing a single-cell suspension according to the method claimed in claim 6; and (b) growing the cells from the single-cell suspension in a cell culture medium.
Claim 19
Dependent Claim
26 words
A method of establishing a primary tumor cell culture as claimed in claim 17, wherein growing the cells in step (b) further comprises passaging the cells.
Claim 20
Independent Claim
85 words
A method of preparing a single cell suspension of tumor cells, the method comprising the steps of: (a) transferring isolated tumor tissue to a cell culture medium; (b) incubating for 10 to 40 minutes; (c) transferring the tumor tissue to fresh cell culture medium; (d) repeating step c) four to five times; (e) cutting the tumor tissue into small pieces; (f) injecting into the tumor tissue cell culture medium; and (g) flushing cells out of the tumor tissue until a single cell suspension is obtained.
Claim 21
Independent Claim
85 words
A method of preparing a single cell suspension of tumor cells, the method comprising the steps of: (a) transferring isolated tumor tissue to a cell culture medium; (b) incubating for 10 to 40 minutes; (c) transferring the tumor tissue to fresh cell culture medium; (d) repeating step c) four to five times; (e) cutting the tumor tissue into small pieces; (f) injecting into the tumor tissue cell culture medium; and (g) flushing cells out of the tumor tissue until a single cell suspension is obtained.
References Cited (Backward Citations)
US5434132A
Cited
US5273963A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-04-27 | 2001-10-27 | 2002-04-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-04-27 | 2005-10-27 | 2006-04-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-04-27 | 2009-10-27 | 2010-04-28 | $7700.00 | Window Closed / Lapsed |