US4645669A
Expired / Lapsed
US
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Culturing and emplacement of differentiated cells in vivo
Assignee: Albert Einstein College of Medicine
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Filing: 1982-10-04
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Granted: 1987-02-24
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Expires: 2002-10-04
Abstract
Abstract A method and culture solution are disclosed which enable in vivo emplacement of differentiated cells derived from in vitro culturing of differentiated cells, with significant retention of their differentiated character. In an alternative embodiment, in vivo culturing of differentiated cells is contemplated. Through the use of extracellular 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 vivo or 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, and 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
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 15:05
CPC Classifications
A; A61; A61L; A61L27/00; A61L27/36; A61L27/38; A61F; A61F2/00; A61F2/02; A61F2/28; C; C12; C12N; C12N5/00; C12N5/0068; C12N2533/00; C12N2533/90
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
44 words
A method for providing an improved in vivo site for promoting the growth of site-specific differentiated cells, comprising providing site-specific connective tissue-derived fibers comprising collagens, non-collagenous proteins and carbohydrates as a substrate for in vivo cell culture of site-specific differentiated cells at the site.
Claim 2
Independent Claim
44 words
A method for providing an improved in vivo site for promoting the growth of site-specific differentiated cells, comprising providing site-specific connective tissue-derived fibers comprising collagens, non-collagenous proteins and carbohydrates as a substrate for in vivo cell culture of site-specific differentiated cells at the site.
Claim 3
Dependent Claim
14 words
The method of claim 1 in which the site-specific differentiated cells are mammalian cells.
Claim 4
Dependent Claim
14 words
The method of claim 1 in which the site-specific differentiated cells are epithelial cells.
Claim 5
Dependent Claim
23 words
The method of claim 1 in which the site-specific differentiated cells are body cells that are normally in contact with basal lamina material.
Claim 6
Dependent Claim
17 words
The method of claim 35 in which the site-specific connective tissue-derived fibers are isolated from organ tissue.
Claim 7
Independent Claim
87 words
A method for the in vivo culturing of site-specific differentiated cells which comprises selecting an in vivo site to be provided with site-specific differentiated cells, preparing said site for the receipt of culture medium, and disposing a quantity of culture medium comprising site-specific connective tissue-derived fibers comprising collagens, non-collagenous proteins and carbohydrates as a substrate for the in situ culturing of site-specific differentiated cells at said in vivo site, and applying differentiated cells to the culture medium whereby in situ growth of site-specific differentiated cells takes place.
Claim 8
Independent Claim
87 words
A method for the in vivo culturing of site-specific differentiated cells which comprises selecting an in vivo site to be provided with site-specific differentiated cells, preparing said site for the receipt of culture medium, and disposing a quantity of culture medium comprising site-specific connective tissue-derived fibers comprising collagens, non-collagenous proteins and carbohydrates as a substrate for the in situ culturing of site-specific differentiated cells at said in vivo site, and applying differentiated cells to the culture medium whereby in situ growth of site-specific differentiated cells takes place.
Claim 9
Dependent Claim
31 words
The method of claim 6 in which the resultant in situ growth of the culture of site-specific differentiated cells in vivo is bound to cells originally at the in vivo site.
Claim 10
Dependent Claim
14 words
The method of claim 6 in which the site-specific differentiated cells are mammalian cells.
Claim 11
Dependent Claim
14 words
The method of claim 6 in which the site-specific differentiated cells are epithelial cells.
Claim 12
Dependent Claim
23 words
The method of claim 6 in which the site-specific differentiated cells are body cells that are normally in contact with basal lamina material.
Claim 13
Dependent Claim
17 words
The method of claim 6 in which the site-specific connective tissue-derived fibers are isolated from organ tissue.
Claim 14
Independent Claim
107 words
A method for the emplacement at an in vivo site of site-specific differentiated cells which comprises providing an initial supply of site-specific differentiated cells, disposing said initial supply in an in vitro culture medium comprising site-specific collagens, non-collagenous proteins and carbohydrates as a substrate for the in vitro cell culture, whereby in vitro culturing of said site-specific differentiated cells takes place and the cell culture remains substantially differentiated, and transferring and transplanting the resulting growth of the site-specific differentiated cell culture to said in vivo site to be provided with site-specific differentiated cells, so that the supply of differentiated cells at said in vivo site is increased.
Claim 15
Independent Claim
107 words
A method for the emplacement at an in vivo site of site-specific differentiated cells which comprises providing an initial supply of site-specific differentiated cells, disposing said initial supply in an in vitro culture medium comprising site-specific collagens, non-collagenous proteins and carbohydrates as a substrate for the in vitro cell culture, whereby in vitro culturing of said site-specific differentiated cells takes place and the cell culture remains substantially differentiated, and transferring and transplanting the resulting growth of the site-specific differentiated cell culture to said in vivo site to be provided with site-specific differentiated cells, so that the supply of differentiated cells at said in vivo site is increased.
Claim 16
Dependent Claim
25 words
The method of claim 12, wherein the temperature at which the in vitro culturing method is conducted is optimal for the cellular growth in vitro.
Claim 17
Dependent Claim
12 words
The method of claim 12, wherein the site-specific differentiated cells are mammalian.
Claim 18
Dependent Claim
13 words
The method of claim 12, wherein the site-specific differentiated cells are non-mammalian cells.
Claim 19
Dependent Claim
13 words
The method of claim 12, wherein the site-specified differentiated cells are epithelial cells.
Claim 20
Dependent Claim
22 words
The method of claim 12, wherein the site-specific differentiated cells are body cells that are normally in contact with basal lamina material.
Claim 21
Dependent Claim
16 words
The method of claim 12, wherein the site-specific connective tissue-derived fibers are isolated from organ tissue.
Claim 22
Dependent Claim
28 words
The method of claim 12, further comprising placing the site-specific connective tissue-derived fibers onto culture solid-state supports in vitro, and allowing said fibers to cover said solid-state supports.
Claim 23
Dependent Claim
20 words
The method of claim 19, further comprising sterilizing said solid-state supports in vitro, after adding the site-specific connective tissue-derived fibers.
Claim 24
Dependent Claim
21 words
The method of claim 20, further comprising adding the site-specific differentiated cells to the tissue culture plates in vitro after sterilization.
Claim 25
Dependent Claim
18 words
The method of claim 20, wherein the site-specific connective tissue-derived fibers attach to the solid-state support in vitro.
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) | 1990-02-23 | 1990-08-25 | 1991-02-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-02-23 | 1994-08-25 | 1995-02-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-02-23 | 1998-08-25 | 1999-02-24 | $7700.00 | Window Closed / Lapsed |