Bioreactor device with application as a bioartificial liver

A bioreactor apparatus for maintaining cells in vitro over a sustained period of time for continuous production of a desired cell product, said cells being selected from the group consisting of animal cells, genetically altered animal cells and mixtures thereof, said apparatus comprising:

• (a) a housing having a first and a second base plate, the first base plate having first and second fluid inlet means for supplying nutrient medium, and the second base plate having first and second fluid outlet means for removing cell waste products;
• (b) at least one cell growth plate having longitudinal windows formed therethrough and extending substantially the distance from the second fluid inlet means to the second fluid outlet means;
• (c) at least one nutrient medium plate having windows formed therethrough and extending substantially the distance from the first fluid inlet means to the first fluid outlet means;
• (d) at least one selectively permeable biocompatible membrane permeable to the passage of cell nutrient and cell waste while substantially impermeable to the passage of said cells and the desired cell product;
• (e) a substantially insoluble, biocompatible matrix means formed in situ from a solution of matrix precursor in said growth plate for entrapping said cells;
• with the base plates, growth plate, medium plate and membrane being assembled, such that each growth plate alternates with each nutrient medium plate, with a membrane separating each medium plate from each growth plate,
• thereby each of the windows of each growth plate forms, with the adjacent membrane, a cell chamber in flow communication between the second fluid inlet means and the second fluid flow outlet means, and thereby each of the windows of each nutrient medium plate forms, with the adjacent membrane, a nutrient chamber in flow communication between the first fluid inlet means and the first fluid outlet means, and
• wherein said matrix means is contracted within each said cell chamber so as to form a cell-biocompatible matrix zone and a liquid zone for accumulating the desired cell product, said cells causing a significant degree of the contraction of said matrix means so that at most 90% of each said cell chamber is occupied by said matrix means and at least 10% of each said cell chamber is unobstructed to form said liquid zone.

A hollow fiber bioreactor apparatus for maintaining cells in vitro over a sustained period of time for continuous production of a desired cell product, said cells being selected from the group consisting of animal cells, genetically altered animal cells and mixtures thereof, said apparatus comprising:

• (a) housing means having spaced end portions and defining a chamber therebetween;
• (b) at least one selectively permeable hollow fiber substantially permeable to the passage of nutrients and cell waste products while impermeable to the passage of said cells and the desired cell product, said chamber being divided by the wall of said hollow fiber into an intracapillary space within said hollow fiber and an extracapillary space outside said hollow fiber, said intracapillary space and said extracapillary space communicating with each other only through the wall of said hollow fiber;
• (c) a substantially insoluble, biocompatible matrix means formed in situ within said intracapillary space for entrapping said cells, said matrix means being contracted within said intracapillary space so as to form a cell-biocompatible matrix zone and a liquid zone for accumulating the desired cell product, said cells causing a significant degree of the contraction of said matrix means so that at most 90% of said intracapillary space is occupied by said matrix means and at least 10% of said intracapillary space is unobstructed to form said liquid zone;
• (d) means in flow communication with said intracapillary space for introducing said cells in admixture with a matrix precursor and for withdrawing the desired cell product;
• and
• (e) means in flow communication with said extracapillary space for introducing cell nutrient for maintaining said cells and withdrawing expended cell nutrient and cell waste from said extracapillary space.

A hollow fiber bioreactor apparatus for maintaining cells in vitro over a sustained period of time for continuous production of at least a first and a second desired cell product, said cells being selected from the group consisting of animal cells, genetically altered animal cells and mixtures thereof, said apparatus comprising:

• (a) housing means having spaced end portions and defining a chamber therebetween;
• (b) a first selectively permeable hollow fiber residing within said chamber, said first hollow fiber being substantially permeable to the passage of cell nutrient and cell waste and to the second desired cell product while impermeable to the passage of said cells and the first desired cell product;
• (c) a second selectively permeable hollow fiber concentric to said first hollow fiber, said second hollow fiber being substantially permeable to the passage of cell nutrient and cell waste while impermeable to the passage of the first and second desired cell products, wherein said chamber is divided by said first and second fibers into three zones, a first zone within the intracapillary space of said first hollow fiber, a second zone within the intracapillary space intermediate said first hollow fiber and said second hollow fiber, and a third zone within the extracapillary space intermediate said second hollow fiber and said housing means;
• (d) a substantially insoluble, biocompatible matrix means formed in situ from a suspension of matrix precursor within said first zone for entrapping said cells, said matrix means being contracted within said first zone so as to form a cell-biocompatible matrix phase and a liquid phase for accumulating the first and second desired cell products, said cells causing a significant degree of the contraction of said matrix means so that at most 90% of said first zone is occupied by said matrix means and at least 10% of said first zone is unobstructed to form said liquid phase;
• (e) means in flow communication with said first zone for introducing said cells in admixture with a matrix precursor and for withdrawing the first and second desired cell product;
• (f) means in flow communication with said second zone for withdrawing the second desired cell product;
• and
• (g) means in flow communication with said third zone for introducing cell nutrient for maintaining said cells and withdrawing expended cell nutrient and cell waste from said third zone.

A hollow fiber bioreactor apparatus for maintaining cells in vitro over a sustained period of time for continuous production of at least a first and a second desired cell product, said cells being selected from the group consisting of animal cells, genetically altered animal cells and mixtures thereof, said apparatus comprising:

• (a) housing means having spaced end portions and defining a chamber therebetween;
• (b) a first selectively permeable hollow fiber residing within said chamber, said first hollow fiber being substantially permeable to the passage of cell nutrient, cell waste and the first and second desired cell products while impermeable to the passage of said cells;
• (c) a second selectively permeable hollow fiber concentric to said first hollow fiber, said second hollow fiber being substantially permeable to the passage of cell nutrient and cell waste of said cells and to the second desired cell product while impermeable to the first desired cell product;
• (d) a third selectively permeable hollow fiber concentric to said second hollow fiber, said third hollow fiber being substantially permeable to the passage of cell nutrient and cell waste while impermeable to the first and second desired cell products, said chamber being divided by said first, second and third hollow fibers into four zones, a first zone within the intracapillary space of said first hollow fiber, a second zone within the intracapillary space intermediate said first hollow fiber and said second hollow fiber, a third zone within the intracapillary space intermediate said third hollow fiber and said second hollow fiber and a fourth zone within the extracapillary space intermediate said third hollow fiber and said housing means;
• (e) a substantially insoluble, biocompatible matrix means formed in situ from a suspension of matrix precursor within said first zone for entrapping said cells, said matrix means being contracted within said first zone so as to form a cell-biocompatible matrix phase and a liquid phase for accumulating the first and second desired cell products, said cells causing a significant degree of the contraction of said matrix means so that at most 90% of said first zone is occupied by said matrix means and at least 10% of said first zone is unobstructed to form said liquid phase;
• (f) means in flow communication with said first zone for introducing said cells in admixture with a matrix precursor and for withdrawing the first and second desired cell products;
• (g) means in flow communication with said third zone for withdrawing the second desired cell product;
• and
• (h) means in flow communication with said fourth zone for introducing cell nutrient for maintaining said cells and withdrawing expended cell nutrient and cell waste from said fourth zone.

A hollow fiber cell culture bioreactor comprising a housing having first inlet and outlet ports defining a fluid flow cavity therebetween, said cavity also enclosing a plurality of selectively permeable hollow fibers in fluid flow communication with second inlet and outlet ports in said housing, each of said hollow fibers having a lumen and a selectively permeable wall, such that communication between the cavity and the lumen of each of the hollow fibers is exclusively through the selectively permeable wall, and each of said hollow fibers being substantially permeable to the passage of cell nutrient and waste while impermeable to the passage of cells and a desired cell product, said bioreactor having three separate cell culture zones:

• a first zone comprising an insoluble fibrous and porous cell gel matrix contracted within the lumen of at least one of the hollow fibers;
• a second zone within the lumen of the at least one of the hollow fibers and around the contracted cell gel matrix;
• and
• a third zone within the fluid flow cavity around the hollow fibers;
• wherein said cells contained in said cell gel matrix cause a significant degree of the contraction of said cell gel matrix so that at most 90% of the lumen of the at least one of the hollow fibers is occupied by said cell gel matrix and at least 10% of the lumen of the at least one of the hollow fibers is unobstructed to form said second zone.

A bioreactor device for retaining cultured liver cells in vitro over a sustained period of time, comprising:

• (a) a housing having a first and a second base plate, the first base plate having first and second fluid inlet means for supplying nutrient medium, and the second base plate having first and second fluid outlet means for removing cell waste products;
• (b) at least one cell growth plate having windows formed therethrough and extending substantially the distance from the second fluid inlet means to the second fluid outlet means;
• (c) at least one nutrient medium plate having windows formed therethrough and extending substantially the distance from the first fluid inlet means to the first fluid outlet means;
• (d) at least one selectively permeable membrane permeable to the passage of cell nutrient and cell waste while substantially impermeable to liver cells, the base plates, growth plate, medium plate and membrane assembled together such that each growth plate alternates with each medium plate with said membrane separating each medium plate from each growth plate;
• with each of the windows of each growth plate forming, with the adjacent membrane, a cell chamber for liver cells and in flow communication between the second fluid inlet means and the second fluid outlet means, and with each of the windows of each medium plate forming with the adjacent membrane a nutrient chamber in flow communication between said first fluid inlet means and the first fluid outlet means, said bioreactor device further comprising:
• a substantially insoluble, biocompatible matrix means formed in situ in each said cell chamber and for entrapping the liver cells, and
• wherein said matrix means is contracted within each said cell chamber so as to form a cell-biocompatible matrix zone and a liquid zone, said liver cells causing a significant degree of the contraction of said matrix means so that at most 90% of each said cell chamber is occupied by said matrix means and at least 10% of each said cell chamber is unobstructed to form said liquid zone.

A bioreactor apparatus for maintaining cells in vitro over a sustained period of time for continuous or periodic production of a desired cell product, said cells being selected from the group of animal cells, genetically altered animal cells and mixtures thereof, said apparatus comprising:

• (a) a housing having a proximal end and a distal end;
• (b) at least one selectively permeable membrane within said housing extending from said proximal end to said distal end of said housing, said membrane dividing the interior of said housing into a cell chamber and a chamber for cell nutrient and cell waste, said membrane selectively allowing cell nutrient and cell waste to cross between said chambers but retaining the desired cell product within the cell chamber;
• (c) said cell chamber being subdivided into two phases comprising a liquid zone for accumulating the desired cell product, and a substantially insoluble, biocompatible matrix, the matrix comprising a fibrous gel formed in situ from a solution of matrix precursors in the cell chamber, and cells being entrapped within the gel during the formation thereof;
• and
• (d) means communicating with said chamber for cell nutrient and cell waste for supplying nutrient medium for maintaining said cells and for withdrawing expended cell nutrient and cell waste.