EP0103081A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Method of growing cells in vitro
Assignee: Strategic Diagnostics Inc
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Filing: 1983-06-18
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Granted: —
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Expires: 2003-06-18
Abstract
Abstract An improved method and apparatus is disclosed for adding fluids to, or removing fluids from, a body part or organ of an animal which minimizes animal trauma and permits rapid, easy, repeated fluid or low viscosity gel transfers. The apparatus includes a tubular fluid-conveying element adapted for fixed connection to the animal, along with a flexible fluid conduit connected between the element and a specific internal body part or organ, such as a cow's uterus. In use, a syringe is employed to introduce or remove fluids from the body part, through the tubular element and connected conduit. The invention is especially adapted for introducing and recovering cell lines producing monoclonal antibodies or other biologically active products in large mammals, and facilitates monitoring of antibody production as well as administration of nutrients to enhance cell line growth. In preferred monoclonal antibody production procedures, the cells are initially educated through in vitro contact with ambient fluid of the selected organ (e.g., allantoic fluid from a pregnant host), whereupon the educated cells are inoculated and allowed to incubate in the host. It has been found particularly advantageous to employ the allantois of a pregnant mammal for antibody production, and some or all of the cell incubation period therein should occur after the onset of immunocompetency of the fetus.
Inventors
- 👤 James S. Cullor
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 13:27
CPC Classifications
A; A61; A61M; A61M39/00; A61M39/02; A61M39/0247; A61D; A61D7/00; C; C12; C12N; C12N5/00; C12N5/10; C12N5/12; C12N5/16; C12N5/163; C12N2502/00; C12N2502/02; C12N2510/00; C12N2510/02
Claims Analysis
Total Claims: 43
Claim 1
Independent Claim
242 words
Apparatus for selectively withdrawing fluids from, or adding fluids to, and animal's body, said apparatus comprising: an assembly including a tubular, fluid-conveying element, and means operably coupled with said element for normally blocking fluid flow therethrough and being openahle for selectively permitting fluid flow through said element; means for operatively securing said assembly to said animal's body with the outer end of said element being adjacent the exterior surface of the animal's skin; an elongated, flexible, fluid-conveying conduit having an inner end and an outer end, said outer end being operably coupled and in fluid flow communication with said element when said assembly is operatively secured to said animal, said inner conduit end being configured for insertion through the wall of a specific organ or the like of said animal, with the innermost portion of the flexible conduit lying within the organ or the like, said conduit being of a length greater than the shortest distance between said element and the point of insertion thereof through the wall of said organ or the like for, in conjunction with the flexibility of the conduit, permitting and accommodating natural movement of said organ or the like over time; and means for physically attaching and interlocking said inner end of said conduit to said wall of said organ or the like with said innermost portion lying within the organ or the like, including structure for preventing leakage of fluid from said organ or the like.
Claim 2
Dependent Claim
242 words
Apparatus for selectively withdrawing fluids from, or adding fluids to, and animal's body, said apparatus comprising: an assembly including a tubular, fluid-conveying element, and means operably coupled with said element for normally blocking fluid flow therethrough and being openahle for selectively permitting fluid flow through said element; means for operatively securing said assembly to said animal's body with the outer end of said element being adjacent the exterior surface of the animal's skin; an elongated, flexible, fluid-conveying conduit having an inner end and an outer end, said outer end being operably coupled and in fluid flow communication with said element when said assembly is operatively secured to said animal, said inner conduit end being configured for insertion through the wall of a specific organ or the like of said animal, with the innermost portion of the flexible conduit lying within the organ or the like, said conduit being of a length greater than the shortest distance between said element and the point of insertion thereof through the wall of said organ or the like for, in conjunction with the flexibility of the conduit, permitting and accommodating natural movement of said organ or the like over time; and means for physically attaching and interlocking said inner end of said conduit to said wall of said organ or the like with said innermost portion lying within the organ or the like, including structure for preventing leakage of fluid from said organ or the like.
Claim 3
Dependent Claim
15 words
Apparatus as set forth in Claim 1, including selectively operable valve means within said assembly.
Claim 4
Dependent Claim
85 words
Apparatus as set forth in Claim 2, said valve means including: a flow blocking plug located within said element; structure defining a wall surface adjacent said plug and complemental with the plug; means for biasing said plug against said wall surface for blocking fluid flow past the plug; and means engageable with the plug for selectively shifting the plug away from said wall surface, and for permitting fluid flow through said element and past said plug when the plug is in said shifted away position.
Claim 5
Dependent Claim
19 words
Apparatus as set forth in Claim 1, said innermost portion of said conduit including a plurality of apertures therethrough.
Claim 6
Dependent Claim
24 words
Apparatus as set forth in Claim 1, said attaching and interlocking means comprising a support collar disposed about said conduit intermediate the ends thereof.
Claim 7
Dependent Claim
25 words
Apparatus as set forth in Claim 5, said support collar being suturable for attachment and interlocking to said wall of said organ or the like.
Claim 8
Dependent Claim
11 words
Apparatus as set forth in Claim 5, said collar being perforate.
Claim 9
Dependent Claim
11 words
Apparatus as set forth in Claim 5, said collar being flexible.
Claim 10
Dependent Claim
21 words
Apparatus as set forth in Claim 5, said collar extending at least about one-quarter of an inch outwardly from said conduit.
Claim 11
Dependent Claim
24 words
Apparatus as set forth in Claim 1, said innermost portion of said conduit being of a different diameter than the remainder of said conduit.
Claim 12
Dependent Claim
22 words
Apparatus as set forth in claim 10, said innermost portion having a diameter greater than that of the remainder of said conduit.
Claim 13
Dependent Claim
26 words
Apparatus as set forth in clam 1, said conduit comprising, for at least a segment of the length thereof, a pair of telescopically interfitted tubular components.
Claim 14
Dependent Claim
26 words
Apparatus as set forth in clam 1, said conduit comprising, for at least a segment of the length thereof, a pair of telescopically interfitted tubular components.
Claim 15
Dependent Claim
28 words
Apparatus as set. forth in Claim 1, said assembly-securing means comprising an outer tubular component receiving said tubular element, and an external screw thread about said tubular component.
Claim 16
Independent Claim
225 words
A method of periodically and selectively removing fluids from, or adding fluids to, a specific organ or the like of a living animal said method comprising the steps of: providing a tubular, fluid-conveying element; operatively securing said element to said animal's body with the outer end of the element adjacent the exterior surface of the animal's skin; operatively coupling an elongated, flexible, fluid-conveying conduit to, respectively, said element and said organ or the like, said coupling step including the steps of making an incision in the wall of said organ or the like and inserting the inner end of said conduit into said organ or the like until the inntermost portion of the cnduit lies within the organ or the like; said conduit being of a length greater than the shortest distance between said element and said incision for, in conjunction with the flexibility of the conduit, permitting and accommodating natural movement of said organ or the like over time; physically attaching and interlocking said inner end of aid conduit to said wall of said organ 'or the like, with said innermost portion lying within the organ or the like, for preventing leakage of fluid from said organ or the like; and periodically and selectively withdrawing fluid from, or adding fluid to, said organ or the like by conveying fluid through said conduit and element.
Claim 17
Independent Claim
225 words
A method of periodically and selectively removing fluids from, or adding fluids to, a specific organ or the like of a living animal said method comprising the steps of: providing a tubular, fluid-conveying element; operatively securing said element to said animal's body with the outer end of the element adjacent the exterior surface of the animal's skin; operatively coupling an elongated, flexible, fluid-conveying conduit to, respectively, said element and said organ or the like, said coupling step including the steps of making an incision in the wall of said organ or the like and inserting the inner end of said conduit into said organ or the like until the inntermost portion of the cnduit lies within the organ or the like; said conduit being of a length greater than the shortest distance between said element and said incision for, in conjunction with the flexibility of the conduit, permitting and accommodating natural movement of said organ or the like over time; physically attaching and interlocking said inner end of aid conduit to said wall of said organ 'or the like, with said innermost portion lying within the organ or the like, for preventing leakage of fluid from said organ or the like; and periodically and selectively withdrawing fluid from, or adding fluid to, said organ or the like by conveying fluid through said conduit and element.
Claim 18
Dependent Claim
13 words
The method of Claim 14, said organ or the like being a uterus.
Claim 19
Dependent Claim
19 words
The method of Claim 15, including the step of inserting said inner end of the conduit into the allantois.
Claim 20
Independent Claim
76 words
A method of growing cells comprising the steps of: selecting a body cavity of a living animal as a situs for cell growth; obtaining ambient fluid characteristic of said body cavity; educating said cells by contacting the cells with said ambient fluid, in vitro; innoculating said educated cells into said selected cavity containing said ambient fluid; allowing said cells to multiply in said ambient fluid; and harversting said cells or the secretory products of said cells.
Claim 21
Independent Claim
76 words
A method of growing cells comprising the steps of: selecting a body cavity of a living animal as a situs for cell growth; obtaining ambient fluid characteristic of said body cavity; educating said cells by contacting the cells with said ambient fluid, in vitro; innoculating said educated cells into said selected cavity containing said ambient fluid; allowing said cells to multiply in said ambient fluid; and harversting said cells or the secretory products of said cells.
Claim 22
Dependent Claim
28 words
The method of Claim 17, said educating step comprising the steps of repeatedly contacting said cells with respective quantities of said ambient fluid over a period of time.
Claim 23
Dependent Claim
15 words
The method of Claim 18, said quantities increasing in volume over said period of time.
Claim 24
Dependent Claim
18 words
The method of Claim 17, said ambient fluid being amnionic fluid from the uterus of a pregnant host.
Claim 25
Dependent Claim
45 words
The method of Claim 17, said cavity being a uterus of a pregnant host, said ambient fluid having uterine fluid of said host, said host being an animal wherein the animal's placenta serves as a barrier to the passage of maternal antibodies into the uterus.
Claim 26
Dependent Claim
24 words
The method of Claim 21, said host being selected from the group consisting of sheep, horses, pigs, and cows, said cavity being the uterus.
Claim 27
Dependent Claim
21 words
The method of Claim 17, said cells being maintained at a temperature of about 38.3-39 degrees centigrade during said educating step.
Claim 28
Dependent Claim
13 words
The method of Claim 17, said ambient fluid being derived from said host.
Claim 29
Dependent Claim
19 words
The method of Claim 17, said ambient fluid being obtained from the same type of animal as said host.
Claim 30
Dependent Claim
56 words
The method of Claim 17, said educating step comprising the steps of: contacting said cells with a fluid containing from about 5 to 10% by volume of said ambient fluid,'and allowing said cells to cultivate therein; and thereafter contacting said cells in a stepwise manner with respective fluid samples containing increasing quantities of said ambient fluid.
Claim 31
Dependent Claim
27 words
The method of Claim 17, said cells being selected from the group consisting of hybridoma cells or any cell line producing a biologically active protein or product.
Claim 32
Independent Claim
35 words
A method of in vitro cell growth comprising contacting cells to be grown with the uterine fluid of a pregnant animal under conditions whereby said cells will multiply, and allowing said cells to grow therein.
Claim 33
Independent Claim
35 words
A method of in vitro cell growth comprising contacting cells to be grown with the uterine fluid of a pregnant animal under conditions whereby said cells will multiply, and allowing said cells to grow therein.
Claim 34
Dependent Claim
11 words
The method of Claim 27, said uterine fluid being allantoic fluid.
Claim 35
Dependent Claim
28 words
The method of Claim 27, including the step of repeatedly contacting said cells with respective fluids containing increasing concentration of said uterine fluid over a period of time.
Claim 36
Independent Claim
60 words
A method of growing cells, comprising the steps of: inoculating said cells into the allantois of a pregnant host; causing said cells to multiply in said allantois over a period of time, at least a portion of said time period being after the onset of imunocompetency of the host's fetus; and thereafter harvesting said cells or the secretory products thereof..
Claim 37
Independent Claim
60 words
A method of growing cells, comprising the steps of: inoculating said cells into the allantois of a pregnant host; causing said cells to multiply in said allantois over a period of time, at least a portion of said time period being after the onset of imunocompetency of the host's fetus; and thereafter harvesting said cells or the secretory products thereof..
Claim 38
Dependent Claim
19 words
The method of Claim 31, said host being selected from the group consisting of sheep, horses, pigs, and cows.
Claim 39
Dependent Claim
37 words
The method of Claim 31, including the step of, prior to said inoculation, contacting said cells in vitro with a quantity of allantoic fluid from the host or from the same type of animal as the host.
Claim 40
Dependent Claim
29 words
The method of Claim 31, said in vitro contacting step comprising the steps of repeatedly contacting said cells with respective fluid samples containing increasing concentrations of said allantoic fluid.
Claim 41
Dependent Claim
18 words
The method of Claim 31, said inoculation being carried out after the onset of imunocompetency of said fetus.
Claim 42
Dependent Claim
24 words
The method of Claim 31, including the step of adding nutrients and/or growth factors for said cells to the allantoic during said time period.
Claim 43
Dependent Claim
10 words
The method of Claim 31, said host being a cow.
References Cited (Backward Citations)
No citation records indexed.
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