US4352887A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method and article for culturing differentiated cells
Assignee: Albert Einstein College of Medicine
•
Filing: 1979-10-29
•
Granted: 1982-10-05
•
Expires: 1999-10-29
Abstract
Abstract A method, solid-state culture support and culture solution are disclosed which enables successful in vitro culturing of differentiated cells, with significant retention of their differentiated character. Through the use of extra-cellular matrix fibers, specifically derived from connective tissue, as culture substrates, the method also discloses the isolation of the connective tissue fibers and their preparation as a culture substrate. This method provides significantly higher survival and attachment rates, and often significantly improved growth properties for in vitro cultures of differentiated cells, especially epithelial, over the current methods for culturing these cells. This method also significantly enables certain differentiated cells to retain much of their normal enzymatic activities. Furthermore, this method enables certain differentiated cells to retain to a high degree, their ability to secrete substances, such as hormones.
Inventors
- 👤 Lola C. M. Reid
- 👤 Marcos Rojkind
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 15:27
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0068; C12N2533/00; C12N2533/90; Y; Y10; Y10S; Y10S623/00; Y10S623/915
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
38 words
A method for in vitro culturing of differentiated cells comprising providing connective tissue-derived fibers comprising collagens, non-collagenous proteins and carbohydrates as a substrate for an in vitro differentiated cell culture whereby the differentiated cell culture remains substantially differentiated.
Claim 2
Independent Claim
38 words
A method for in vitro culturing of differentiated cells comprising providing connective tissue-derived fibers comprising collagens, non-collagenous proteins and carbohydrates as a substrate for an in vitro differentiated cell culture whereby the differentiated cell culture remains substantially differentiated.
Claim 3
Dependent Claim
21 words
The method of claim 1, wherein the temperature at which the method is conducted is between 0° C. and 100° C.
Claim 4
Dependent Claim
24 words
The method of claim 2, wherein the temperature at which the method is conducted for mammalian cells is between 29° C. and 42° C.
Claim 5
Dependent Claim
22 words
The method of claim 2, wherein the temperature at which the method is conducted is optimal for the cellular growth in vitro.
Claim 6
Dependent Claim
34 words
The method of claim 1, wherein the differentiated cells in culture remain differentiated for at least about one month on the connective tissue--derived fibrous substrate so long as nutrients for the cells are provided.
Claim 7
Dependent Claim
11 words
The method of claim 1, wherein the differentiated cells are mammalian.
Claim 8
Dependent Claim
12 words
The method of claim 1, wherein the differentiated cells are non-mammalian cells.
Claim 9
Dependent Claim
12 words
The method of claim 1, wherein the differentiated cells are epithelial cells.
Claim 10
Dependent Claim
21 words
The method of claim 1, wherein the differentiated cells are body cells that are normally in contact with basal lamina material.
Claim 11
Dependent Claim
12 words
The method of claim 1, wherein the differentiated cells are normally-growing cells.
Claim 12
Dependent Claim
17 words
The method of claim 1, wherein in the differentiated cells survive in vitro in a differentiated state.
Claim 13
Dependent Claim
12 words
The method of claim 1, wherein the differentiated cells are malignantly-growing cells.
Claim 14
Independent Claim
15 words
The method of claim, 1 wherein the connective tissue-derived fibers are isolated from organ tissue.
Claim 15
Independent Claim
25 words
The method of claim, 1 further comprising placing the connective tissue-derived fibers onto culture solid-state supports and allowing said fibers to cover said solid-state supports.
Claim 16
Dependent Claim
17 words
The method of claim 14, further comprising sterilizing said solid-state supports after adding the connective tissues-derived fibers.
Claim 17
Dependent Claim
18 words
The method of claim 15, further comprising adding the differentiated cells to the tissue culture plates after sterilization.
Claim 18
Dependent Claim
15 words
The method of claim 14, wherein the connective tissue-derived fibers attach to the solid-state support.
Claim 19
Dependent Claim
46 words
The method of claim 1, wherein certain differentiated cells in culture exhibit a growth rate of at least 300% as measured by the rate of the number of differentiated cells after one month to the number of differentiated cells that attached day 1 of the culture.
Claim 20
Dependent Claim
45 words
The method of claim 1, wherein certain differentiated cells in culture exhibit a growth rate of about 1000% as measured by the ratio of the number of differentiated cells after one month to the number of differentiated cells that attached day 1 of the culture.
Claim 21
Dependent Claim
38 words
The method of claim 1, wherein the time period the differentiated cells in culture remain essentially differentiated is at least about one month and is proportional to the presence of the fibrous substrate and nutrients for the cells.
Claim 22
Dependent Claim
21 words
The method of claim 1, wherein the attachment efficiency of differentiated cells of the in vitro culture is at least 50%.
Claim 23
Dependent Claim
24 words
The method of claim 1, wherein the attachment efficiency of the differentiated cells of the in vitro culture is up to and including 86%.
Claim 24
Dependent Claim
46 words
A cell culture solid-state support consisting essentially of a base support for containing a culture medium and said cells cultured thereon and a plurality of connective tissue derived fibers disposed within said base support, said connective tissue-derived fibers consisting essentially of collagens, non-collagenous proteins and carbohydrates.
Claim 25
Dependent Claim
46 words
A cell culture solid-state support consisting essentially of a base support for containing a culture medium and said cells cultured thereon and a plurality of connective tissue derived fibers disposed within said base support, said connective tissue-derived fibers consisting essentially of collagens, non-collagenous proteins and carbohydrates.
Claim 26
Dependent Claim
28 words
The cell culture solid-state support of claim 23, further comprising a basal medium supplemented with hormones and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 27
Dependent Claim
28 words
The cell culture solid-state support of claim 23, further comprising a basal medium supplemented with plasma and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 28
Dependent Claim
28 words
The cell culture solid-state support of claim 23, further comprising a basal medium supplemented with serum and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 29
Dependent Claim
30 words
The cell culture solid-state support of claim 23, further comprising a basal medium supplemented with hormones and plasma and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 30
Dependent Claim
30 words
The cell culture solid-state support of claim 23, further comprising a basal medium supplemented with hormones and serum and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 31
Dependent Claim
20 words
The cell culture solid state support of claim 33, wherein said connective tissue-derived fibers are attached to said base support.
Claim 32
Dependent Claim
17 words
The cell culture solid-state support of claim 23, wherein said connective tissue-derived fibers cover said base support.
Claim 33
Dependent Claim
16 words
The cell culture solid-state support of claim 23, said connective tissue-derived fibers being from organ tissue.
Claim 34
Dependent Claim
19 words
The cell culture solid-state support of claim 23, being operable for cell growth between 0° C. and 100° C.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1985-10-04 | 1986-04-05 | 1986-10-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-10-04 | 1990-04-05 | 1990-10-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-10-04 | 1994-04-05 | 1994-10-05 | $7700.00 | Window Closed / Lapsed |