Method for in vitro reproduction and growth of cells in culture medium

Claim 1

A method for the in vitro culturing of animal cells in a culture medium to produce, from an initial viable cell mass of particular cell number, a viable cell mass of larger cell number, comprising the steps of: (a) providing a series of animal cell culture sub-compartments, each sub-compartment comprising a culture space for animal cells and for culture medium, defined by a flexible material permeable to oxygen, said series further comprising regulating means, operable external of the culture space, for variably permitting or precluding liquid communication between any given sub-compartment in said series and at least a next serial sub-compartment in said series; (b) arranging said series of animal cell culture sub-compartments in a controlled gaseous environment, comprising oxygen, suitable for in vitro animal cell culture; (c) introducing culture medium and a viable cell mass of particular cell number into a first culture sub-compartment of said series, isolated by said regulating means from liquid communication with any other sub-compartment of said series; (d) maintaining in said first culture sub-compartment conditions effective to cause said viable cell mass therein to increase in cell number (e) thereafter establishing liquid communication, by said regulating means, between said first culture sub-compartment and a next serial culture sub-compartment of said series provided with at least additional culture medium or culture space, and transferring said viable cell mass of increased cell number, and culture medium, from said first culture sub-compartment to said next serial culture sub-compartment; (f) maintaining in said next serial sub-compartment conditions effective to cause said cell mass therein to further increase in cell number; and (g) withdrawing from said next serial culture sub-compartment said viable cell mass of further increased cell number.

Claim 2

A method for the in vitro culturing of animal cells in a culture medium to produce, from an initial viable cell mass of particular cell number, a viable cell mass of larger cell number, comprising the steps of: (a) providing a series of animal cell culture sub-compartments, each sub-compartment comprising a culture space for animal cells and for culture medium, defined by a flexible material permeable to oxygen, said series further comprising regulating means, operable external of the culture space, for variably permitting or precluding liquid communication between any given sub-compartment in said series and at least a next serial sub-compartment in said series; (b) arranging said series of animal cell culture sub-compartments in a controlled gaseous environment, comprising oxygen, suitable for in vitro animal cell culture; (c) introducing culture medium and a viable cell mass of particular cell number into a first culture sub-compartment of said series, isolated by said regulating means from liquid communication with any other sub-compartment of said series; (d) maintaining in said first culture sub-compartment conditions effective to cause said viable cell mass therein to increase in cell number (e) thereafter establishing liquid communication, by said regulating means, between said first culture sub-compartment and a next serial culture sub-compartment of said series provided with at least additional culture medium or culture space, and transferring said viable cell mass of increased cell number, and culture medium, from said first culture sub-compartment to said next serial culture sub-compartment; (f) maintaining in said next serial sub-compartment conditions effective to cause said cell mass therein to further increase in cell number; and (g) withdrawing from said next serial culture sub-compartment said viable cell mass of further increased cell number.

Claim 11

A method for the in vitro culturing of animal cells in a culture medium to produce, from an initial viable cell mass of particular cell number, a viable cell mass of larger cell number, comprising the steps of: (a) providing an enclosed overall culture compartment of defined surface area and volume comprising a flexible oxygen-permeable material, said overall culture compartment being divided into two or more culture sub-compartments by removal segmenting means displaced about the periphery of the enclosed culture compartment; (b) arranging said overall culture compartment in a controlled gaseous environment, comprising oxygen, suitable for in vitro cell culture; (c) introducing culture medium and a viable cell mass of particular cell number into a first of said sub-compartments; (d) maintaining in said first sub-compartment conditions effective to cause said viable cell mass therein to increase in cell number; (e) thereafter removing said segmenting means between said first sub-compartment and a next serial sub-compartment, whereby there is produced a united larger culture space, comprised of said first sub-compartment and said next serial sub-compartment, for culture of the cells from said first sub-compartment; and (f) maintaining in said united larger culture space conditions effective to cause the cell mass therein to further increase in cell number.

Claim 12

A method for the in vitro culturing of animal cells in a culture medium to produce, from an initial viable cell mass of particular cell number, a viable cell mass of larger cell number, comprising the steps of: (a) providing an enclosed overall culture compartment of defined surface area and volume comprising a flexible oxygen-permeable material, said overall culture compartment being divided into two or more culture sub-compartments by removal segmenting means displaced about the periphery of the enclosed culture compartment; (b) arranging said overall culture compartment in a controlled gaseous environment, comprising oxygen, suitable for in vitro cell culture; (c) introducing culture medium and a viable cell mass of particular cell number into a first of said sub-compartments; (d) maintaining in said first sub-compartment conditions effective to cause said viable cell mass therein to increase in cell number; (e) thereafter removing said segmenting means between said first sub-compartment and a next serial sub-compartment, whereby there is produced a united larger culture space, comprised of said first sub-compartment and said next serial sub-compartment, for culture of the cells from said first sub-compartment; and (f) maintaining in said united larger culture space conditions effective to cause the cell mass therein to further increase in cell number.