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EP0175286A2
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In vitro cell culture system and method
Assignee: Medical College of Wisconsin Research Foundation Inc
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Filing: 1985-09-12
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Granted: —
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Expires: 2005-09-12
Abstract
Abstract In vitro cell culture systems for anchorage-dependent mammalian cells (B) using solid substrates (A) of mitogenic calcium compounds, such as hydroxyapatite and tricalcium phosphate forms of calcium phosphate and calcium carbonate. The calcium solid substrates (A) can be in the form of granules with a particle size of at least 0.050 mm. or in the form of solid bodies and may be either porous or non-porous. Unique features of cells (B) cultured in the stated in vitro cell culture systems include the growth of cells in layers many cells thick, growth of cells thai maintain their phenotype and the ability to culture cells for extended periods of time.
Inventors
- 👤 Herman S. Cheung
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 18:10
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0068; C12N2533/00; C12N2533/10; C12N2533/18
Claims Analysis
Total Claims: 102
Claim 1
Independent Claim
20 words
Glacken, M.W., R.J. Fleishaker, and A.J. Sinskey. 1983. Mammalian cells in culture. Engineering principles and scale-up. Trends in Biotechnology 1:102-108.
Claim 2
Dependent Claim
20 words
Glacken, M.W., R.J. Fleishaker, and A.J. Sinskey. 1983. Mammalian cells in culture. Engineering principles and scale-up. Trends in Biotechnology 1:102-108.
Claim 3
Dependent Claim
12 words
McCoy, T.A., W. Whittle, E. Conway. 1962. Proc. Soc. Exp. Biol. 109:225.
Claim 4
Dependent Claim
12 words
McCoy, T.A., W. Whittle, E. Conway. 1962. Proc. Soc. Exp. Biol. 109:225.
Claim 5
Dependent Claim
12 words
Wiemann, M.C., K. McCarthy, B. Creswick, and P. Calabresi. In Vitro 19:260.
Claim 6
Dependent Claim
12 words
Wiemann, M.C., K. McCarthy, B. Creswick, and P. Calabresi. In Vitro 19:260.
Claim 7
Dependent Claim
18 words
van Wezel, A.L. 1967. Growth of cell strains and primary cells on microcarriers in homogeneous culture. Nature 216:64-65.
Claim 8
Dependent Claim
18 words
van Wezel, A.L. 1967. Growth of cell strains and primary cells on microcarriers in homogeneous culture. Nature 216:64-65.
Claim 9
Dependent Claim
15 words
van Wezel, A.L., in Methods and Applications of Tissue Culture, Academic Press, New York, 1973.
Claim 10
Dependent Claim
15 words
van Wezel, A.L., in Methods and Applications of Tissue Culture, Academic Press, New York, 1973.
Claim 11
Dependent Claim
11 words
Microcarrier Cell Culture, Principles and Methods. Pharmacia Fine Chemicals A.B., 1981.
Claim 12
Dependent Claim
11 words
Microcarrier Cell Culture, Principles and Methods. Pharmacia Fine Chemicals A.B., 1981.
Claim 13
Dependent Claim
13 words
Hubbard, W. 1974. Physiological calcium phosphates as orthopedic biomaterials, Ph.D. Thesis, Marquette University.
Claim 14
Dependent Claim
13 words
Hubbard, W. 1974. Physiological calcium phosphates as orthopedic biomaterials, Ph.D. Thesis, Marquette University.
Claim 15
Dependent Claim
9 words
deGroot: Bioceramics of Calcium Phosphate, CRC Press, Inc., 1983.
Claim 16
Dependent Claim
9 words
deGroot: Bioceramics of Calcium Phosphate, CRC Press, Inc., 1983.
Claim 17
Dependent Claim
15 words
Klagsbrun, M. 1979. Large scale preparation of chondrocytes. Methods Enzymol. Academic Press. New York 58:560-564.
Claim 18
Dependent Claim
15 words
Klagsbrun, M. 1979. Large scale preparation of chondrocytes. Methods Enzymol. Academic Press. New York 58:560-564.
Claim 19
Dependent Claim
20 words
Sokoloff, L.C., J. Malemud and W.T. Green, Jr. 1970. Sulfate incorporation by articular chondrocytes in monolayer culture. Arthritis Rheum 13:118-124
Claim 20
Dependent Claim
20 words
Sokoloff, L.C., J. Malemud and W.T. Green, Jr. 1970. Sulfate incorporation by articular chondrocytes in monolayer culture. Arthritis Rheum 13:118-124
Claim 21
Dependent Claim
23 words
Srivastava, V.M.L., C.J. Malemud, and L. Sokoloff. 1974. Chondroid expression by lapine articular chondrocytes in spinner culture following monolayer growth. Tissue Res. 2:127-136.
Claim 22
Dependent Claim
23 words
Srivastava, V.M.L., C.J. Malemud, and L. Sokoloff. 1974. Chondroid expression by lapine articular chondrocytes in spinner culture following monolayer growth. Tissue Res. 2:127-136.
Claim 23
Dependent Claim
28 words
Trippel, S.B., M.G. Ehrlich, L. Lippiello, and H.J. Mankin. 1980. Characterization of chondrocytes from bovine articular cartilage. I. Metabolic and morphological experimental studies. J. Bone Joint Surge 62-A:816-820.
Claim 24
Dependent Claim
28 words
Trippel, S.B., M.G. Ehrlich, L. Lippiello, and H.J. Mankin. 1980. Characterization of chondrocytes from bovine articular cartilage. I. Metabolic and morphological experimental studies. J. Bone Joint Surge 62-A:816-820.
Claim 25
Dependent Claim
25 words
Oegema, T.R., Jr. and R.C. Thompson, Jr. 1981. Characterization of a hyaluronic acid-dermatan sulfate proteoglycan complex from dedifferentiated human chondrocyte cultures. J. Biol. Chem. 256:1015-1022.
Claim 26
Dependent Claim
25 words
Oegema, T.R., Jr. and R.C. Thompson, Jr. 1981. Characterization of a hyaluronic acid-dermatan sulfate proteoglycan complex from dedifferentiated human chondrocyte cultures. J. Biol. Chem. 256:1015-1022.
Claim 27
Dependent Claim
25 words
Okayama, M., M. Pacifici, and H. Holtzer. 1976. Differences among sulfated proteoglycans synthesized in nonchondrogenic cells, presumptive chondroblasts, and chondroblasts. Proc. Nat. Acad. Sci. 73:3224-3228.
Claim 28
Dependent Claim
25 words
Okayama, M., M. Pacifici, and H. Holtzer. 1976. Differences among sulfated proteoglycans synthesized in nonchondrogenic cells, presumptive chondroblasts, and chondroblasts. Proc. Nat. Acad. Sci. 73:3224-3228.
Claim 29
Dependent Claim
18 words
von der Mark, K. 1980. Immunological studies on collagen type transition in chondrogenesis. Curr. Top. Dev. Biol. 14:199-225.
Claim 30
Dependent Claim
18 words
von der Mark, K. 1980. Immunological studies on collagen type transition in chondrogenesis. Curr. Top. Dev. Biol. 14:199-225.
Claim 31
Dependent Claim
26 words
West, C.M., R. Lanza, J. Rosenbloom, M. Lowe, H. Holtzer, and N. Avdalovic. 1979. Fibronectin alters the phenotypic properties of cultured chick embryo chondroblasts. Cell 17:491-501.
Claim 32
Dependent Claim
26 words
West, C.M., R. Lanza, J. Rosenbloom, M. Lowe, H. Holtzer, and N. Avdalovic. 1979. Fibronectin alters the phenotypic properties of cultured chick embryo chondroblasts. Cell 17:491-501.
Claim 33
Dependent Claim
22 words
Schlitz, J.R., R. Mayne and H. Holtzer, 1973. The synthesis of collagen and glycosaminoglycans by dedifferentiated chondroblasts in culture. Cell Differentiation 1:97-108.
Claim 34
Dependent Claim
22 words
Schlitz, J.R., R. Mayne and H. Holtzer, 1973. The synthesis of collagen and glycosaminoglycans by dedifferentiated chondroblasts in culture. Cell Differentiation 1:97-108.
Claim 35
Dependent Claim
29 words
Mayne, R., M.S. Vail, and E.J. Miller. 1975. Analysis of changes in collagen biosynthesis that occurs when chick chondrocytes are grown in 5-bromo-2'-deoxyuridine. Proc. Natl. Acad. Sci. USA 72:4511-4515.
Claim 36
Dependent Claim
29 words
Mayne, R., M.S. Vail, and E.J. Miller. 1975. Analysis of changes in collagen biosynthesis that occurs when chick chondrocytes are grown in 5-bromo-2'-deoxyuridine. Proc. Natl. Acad. Sci. USA 72:4511-4515.
Claim 37
Dependent Claim
28 words
Mayne, R., M.S. Vail, P.M. Mayne and E.J. Miller. 1976a. The effect of embryo extract on the types of collagen synthesized by cultured chick chondrocytes. Dev. Biol. 54:230-240.
Claim 38
Dependent Claim
28 words
Mayne, R., M.S. Vail, P.M. Mayne and E.J. Miller. 1976a. The effect of embryo extract on the types of collagen synthesized by cultured chick chondrocytes. Dev. Biol. 54:230-240.
Claim 39
Dependent Claim
34 words
Mayne, R., M.S. Vail, P.M. Mayne and E.J. Miller. 1976b. Changes in the type of collagen synthesized as clones of chick chondrocytes grow and eventually lose division capacity. Proc. Natl. Acad. Sci. USA 73:1674-1678.
Claim 40
Dependent Claim
34 words
Mayne, R., M.S. Vail, P.M. Mayne and E.J. Miller. 1976b. Changes in the type of collagen synthesized as clones of chick chondrocytes grow and eventually lose division capacity. Proc. Natl. Acad. Sci. USA 73:1674-1678.
Claim 41
Dependent Claim
27 words
Cheung, H.S., W. Harvey, P.D. Benya, and M.E. Nimni. 1976. New collagen markers of "derepression" synthesized by rabbit articular chondrocytes in culture. Biochem. Biophys. Res. Commun. 68:1371-1378.
Claim 42
Dependent Claim
27 words
Cheung, H.S., W. Harvey, P.D. Benya, and M.E. Nimni. 1976. New collagen markers of "derepression" synthesized by rabbit articular chondrocytes in culture. Biochem. Biophys. Res. Commun. 68:1371-1378.
Claim 43
Dependent Claim
36 words
Benya, P.D., S.R. Padila, and M.E. Nimni. 1977. The progeny of rabbit articular chondrocytes synthesize collagen types I and III and type I trimer, but not type II. Verification by cyanogen bromide peptide analysis. Biochemistry 16:865-872.
Claim 44
Dependent Claim
36 words
Benya, P.D., S.R. Padila, and M.E. Nimni. 1977. The progeny of rabbit articular chondrocytes synthesize collagen types I and III and type I trimer, but not type II. Verification by cyanogen bromide peptide analysis. Biochemistry 16:865-872.
Claim 45
Dependent Claim
20 words
Cheung, H.S. and L.M. Ryan. 1981. A method of determining DNA and chondrocytes content of articular cartilage. Analytical Biochemistry 116:93-97.
Claim 46
Dependent Claim
20 words
Cheung, H.S. and L.M. Ryan. 1981. A method of determining DNA and chondrocytes content of articular cartilage. Analytical Biochemistry 116:93-97.
Claim 47
Dependent Claim
20 words
Miller, E.J. 1971. Isolation and characterization of cyanogen bromide peptides from the 1(II) chain of chick cartilage collagen. Biochemistry 10:3030-3035.
Claim 48
Dependent Claim
20 words
Miller, E.J. 1971. Isolation and characterization of cyanogen bromide peptides from the 1(II) chain of chick cartilage collagen. Biochemistry 10:3030-3035.
Claim 49
Dependent Claim
27 words
Lubin, R.A., G.L. Wong, and D.V. Cohn. 1976. Biochemical characterization with parathyroid hormone and calcitonin of isolated bone cells: Provisional identification of osteoclasts and osteoblasts. Endocrinology 99:526-534.
Claim 50
Dependent Claim
27 words
Lubin, R.A., G.L. Wong, and D.V. Cohn. 1976. Biochemical characterization with parathyroid hormone and calcitonin of isolated bone cells: Provisional identification of osteoclasts and osteoblasts. Endocrinology 99:526-534.
Claim 51
Dependent Claim
28 words
Cheung, H.S., J.T. Nicoloff, M.B. Kamiel, L. Spolter, and M.E. Nimni. 1978. Stimulation of fibroblasts biosynthetic activity by serum of patients with pretibial myxedema. J. Invest. Dermatology. 71:12-17.
Claim 52
Dependent Claim
28 words
Cheung, H.S., J.T. Nicoloff, M.B. Kamiel, L. Spolter, and M.E. Nimni. 1978. Stimulation of fibroblasts biosynthetic activity by serum of patients with pretibial myxedema. J. Invest. Dermatology. 71:12-17.
Claim 53
Dependent Claim
25 words
Cheung, H.S., K.L. Lynch, R.P. Johnson, and B.J. Brewer. 1980. In vitro synthesis of tissue specific type II collagen by healing cartilage. Arthritis Rheum 23:211-219.
Claim 54
Dependent Claim
25 words
Cheung, H.S., K.L. Lynch, R.P. Johnson, and B.J. Brewer. 1980. In vitro synthesis of tissue specific type II collagen by healing cartilage. Arthritis Rheum 23:211-219.
Claim 55
Dependent Claim
28 words
Peterkofsky, B. and R. Diegelmann. 1971. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry 10:988-994.
Claim 56
Dependent Claim
28 words
Peterkofsky, B. and R. Diegelmann. 1971. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry 10:988-994.
Claim 57
Dependent Claim
38 words
Wong, G.L., and D.V. Cohn. 1978. The effect of parathormone on the synthesis of collaginous matrix by isolated bone cells. Proceedings, Mechanisms of Localized Bone Loss. Eds. Horton, Tarpley and Davis. Special Supplement to Calcific Tissue Abstracts 47-59.
Claim 58
Dependent Claim
38 words
Wong, G.L., and D.V. Cohn. 1978. The effect of parathormone on the synthesis of collaginous matrix by isolated bone cells. Proceedings, Mechanisms of Localized Bone Loss. Eds. Horton, Tarpley and Davis. Special Supplement to Calcific Tissue Abstracts 47-59.
Claim 59
Dependent Claim
21 words
Lubin, R.A., and D.V. Cohn. 1976. Effects of parathormone and calcitonin on citrate and hyaluronate metabolism in cultured bone. Endrocinology 98:413-419.
Claim 60
Dependent Claim
21 words
Lubin, R.A., and D.V. Cohn. 1976. Effects of parathormone and calcitonin on citrate and hyaluronate metabolism in cultured bone. Endrocinology 98:413-419.
Claim 61
Dependent Claim
24 words
Cheung, H.S., M.T. Stony, and D.J. McCarty. 1984. Mitogenic effects of hydroxyapatite and calcium pyrophosphate dihydrate crystals on cultured mammalian cells. Arthritis Rheum. 27:668-674.
Claim 62
Dependent Claim
24 words
Cheung, H.S., M.T. Stony, and D.J. McCarty. 1984. Mitogenic effects of hydroxyapatite and calcium pyrophosphate dihydrate crystals on cultured mammalian cells. Arthritis Rheum. 27:668-674.
Claim 63
Dependent Claim
33 words
Cheung, H.S. and D.J. McCarty. 1984. Biological effects of calcium containing crystals on synovio- cytes. In Calcium in Biological System. Eds. R.P. Rubin, G. Weiss, and J.W. Putney, Jr. Plenum Publishing Co., N.Y.
Claim 64
Dependent Claim
33 words
Cheung, H.S. and D.J. McCarty. 1984. Biological effects of calcium containing crystals on synovio- cytes. In Calcium in Biological System. Eds. R.P. Rubin, G. Weiss, and J.W. Putney, Jr. Plenum Publishing Co., N.Y.
Claim 65
Dependent Claim
23 words
Cheung, H.S. and D.J. McCarty. 1983. Calcium containing crystals can substitute for platelet derived growth factor (PDGF) in cell culture. Arthritis Rheum. 26:S60.
Claim 66
Dependent Claim
23 words
Cheung, H.S. and D.J. McCarty. 1983. Calcium containing crystals can substitute for platelet derived growth factor (PDGF) in cell culture. Arthritis Rheum. 26:S60.
Claim 67
Dependent Claim
21 words
Evans, R.W., H.S. Cheung, and D.J. McCarty. 1983. Uptake and dissolution of calcium phosphate crystals by cultured cells. Arthritis Rheum 26:560.
Claim 68
Dependent Claim
21 words
Evans, R.W., H.S. Cheung, and D.J. McCarty. 1983. Uptake and dissolution of calcium phosphate crystals by cultured cells. Arthritis Rheum 26:560.
Claim 69
Dependent Claim
21 words
Evans, R.W., H.S. Cheung, and D.J. McCarty. 1984. Cultured canine synovial cells solubilize 45 Ca labeled hydroxyapatite crystals. Arthritis Rheum. 27:829-832.
Claim 70
Dependent Claim
21 words
Evans, R.W., H.S. Cheung, and D.J. McCarty. 1984. Cultured canine synovial cells solubilize 45 Ca labeled hydroxyapatite crystals. Arthritis Rheum. 27:829-832.
Claim 71
Dependent Claim
21 words
Evans, R.W., H.S. Cheung, and D.J. McCarty. Cultured human monocytes and fibroblasts solubilize calcium phosphate crystals. Calcified Tissue International (In Press).
Claim 72
Dependent Claim
21 words
Evans, R.W., H.S. Cheung, and D.J. McCarty. Cultured human monocytes and fibroblasts solubilize calcium phosphate crystals. Calcified Tissue International (In Press).
Claim 73
Dependent Claim
22 words
Kwong, C.H., and H.S. Cheung. Solubilization of hydroxyapatite crystals by murine bone cells, macrophages and fibroblasts: Effect of PTH. Submitted for publication.
Claim 74
Dependent Claim
22 words
Kwong, C.H., and H.S. Cheung. Solubilization of hydroxyapatite crystals by murine bone cells, macrophages and fibroblasts: Effect of PTH. Submitted for publication.
Claim 75
Dependent Claim
16 words
Cheung, H.S., and D.J. McCarty. Calcium crystal induced mitogenesis: Intracellular dissolution is essential. Submitted for publication.
Claim 76
Dependent Claim
16 words
Cheung, H.S., and D.J. McCarty. Calcium crystal induced mitogenesis: Intracellular dissolution is essential. Submitted for publication.
Claim 77
Dependent Claim
22 words
Dulbecco. R., and J. Elkington. 1975. Induction of growth in resting fibroblastic cell cultures by . Proc. Nat. Acad. Sci. USA 75:1584-1588.
Claim 78
Dependent Claim
22 words
Dulbecco. R., and J. Elkington. 1975. Induction of growth in resting fibroblastic cell cultures by . Proc. Nat. Acad. Sci. USA 75:1584-1588.
Claim 79
Dependent Claim
28 words
Nykforiak, C.J., R.B. Young, T.A. Phillips. 1980. Changes in intracellular distribution during the transition of fibroblasts from the proliferating to the stationary state. Biochem. Biophys. Res. Commun. 3:583-587.
Claim 80
Dependent Claim
28 words
Nykforiak, C.J., R.B. Young, T.A. Phillips. 1980. Changes in intracellular distribution during the transition of fibroblasts from the proliferating to the stationary state. Biochem. Biophys. Res. Commun. 3:583-587.
Claim 81
Dependent Claim
10 words
Akerman, K.E.O. 1982. Transport and cell activation. Medical Biology 60:168-182.
Claim 82
Dependent Claim
10 words
Akerman, K.E.O. 1982. Transport and cell activation. Medical Biology 60:168-182.
Claim 83
Dependent Claim
30 words
Cheung, H.S., P.B. Halverson, and D.J. McCarty. 1982. Release of collagenase, neutral protease and prostaglandins from cultured mammalian synovial cells by hydroxyapatite and calcium pyrophosphate dihydrate crystals. Arthritis Rheum. 24:1338-1344.
Claim 84
Dependent Claim
30 words
Cheung, H.S., P.B. Halverson, and D.J. McCarty. 1982. Release of collagenase, neutral protease and prostaglandins from cultured mammalian synovial cells by hydroxyapatite and calcium pyrophosphate dihydrate crystals. Arthritis Rheum. 24:1338-1344.
Claim 85
Dependent Claim
30 words
Hollemans, M., R.O. Elferink, P.G. DeGroot, A. Strijland, and J.M. Tager. 1981. Accumulation of weak bases in relation to intralysosomal pH in cultured human skin fibroblasts. Biochem. Biophys. Acta 643:140-151..
Claim 86
Dependent Claim
30 words
Hollemans, M., R.O. Elferink, P.G. DeGroot, A. Strijland, and J.M. Tager. 1981. Accumulation of weak bases in relation to intralysosomal pH in cultured human skin fibroblasts. Biochem. Biophys. Acta 643:140-151..
Claim 87
Dependent Claim
30 words
Ohkuma, S., and B. Poole. 1978. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc. Natl. Acad. Sci. USA 75:3327-3331.
Claim 88
Dependent Claim
30 words
Ohkuma, S., and B. Poole. 1978. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc. Natl. Acad. Sci. USA 75:3327-3331.
Claim 89
Dependent Claim
57 words
An in vitro cell culture system for anchorage-dependent mammalian cells (B) of the type including a solid substrate (A) on which the cells are to be grown in an appropriate nutrient growth medium, characterized in that the solid substrate (A) includes a mitogenic calcium compound nontoxic to cells and of a size sufficient to support anchorage-dependent cells.
Claim 90
Dependent Claim
57 words
An in vitro cell culture system for anchorage-dependent mammalian cells (B) of the type including a solid substrate (A) on which the cells are to be grown in an appropriate nutrient growth medium, characterized in that the solid substrate (A) includes a mitogenic calcium compound nontoxic to cells and of a size sufficient to support anchorage-dependent cells.
Claim 91
Dependent Claim
40 words
An in vitro cell culture system according to claim 1 characterized in that the solid substrate (A) is in the form of granules of the calcium compound having a particle size of at least 0.050 millimeters in their largest dimension.
Claim 92
Dependent Claim
31 words
An in vitro cell culture system according to claim 2 characterized in that the granules of the calcium compound have a particle size in the range of 0.050 to 1.0 millimeters.
Claim 93
Dependent Claim
29 words
An in vitro cell culture system according to claim 1 characterized in that the solid substrate (A) is in the form of a solid body of the calcium compound.
Claim 94
Dependent Claim
26 words
An in vitro cell culture system according to claim 4 characterized in that the solid body is formed of sintered porous granules of the calcium compound.
Claim 95
Dependent Claim
34 words
An in vitro cell culture system according to claim 1, 2, 3, 4 or 5 characterized in that the calcium compound is the hydroxyapatite or tricalcium phosphate form of calcium phosphate or calcium carbonate.
Claim 96
Independent Claim
69 words
A method for in vitro culture of anchorage-dependent mammalian cells (B) on the surface of a solid substrate (A) immersed in a nutrient growth medium, characterized by providing a solid substrate (A) including a mitogenic calcium compound that is nontoxic to mammalian cells and of a size sufficient to support anchorage-dependent cells (B), and continuing the cell culture to grow multi-cell thick layers of cells on the solid substrate.
Claim 97
Independent Claim
69 words
A method for in vitro culture of anchorage-dependent mammalian cells (B) on the surface of a solid substrate (A) immersed in a nutrient growth medium, characterized by providing a solid substrate (A) including a mitogenic calcium compound that is nontoxic to mammalian cells and of a size sufficient to support anchorage-dependent cells (B), and continuing the cell culture to grow multi-cell thick layers of cells on the solid substrate.
Claim 98
Dependent Claim
39 words
An in vitro cell culture method according to claim 7 characterized by providing the solid substrate (A) in the form of granules of the calcium compound having a particle size of at least 0.050 millimeters in their largest dimension.
Claim 99
Dependent Claim
31 words
An in vitro cell culture method according to claim 8 characterized in that the granules of the calcium compound have a particle size in the range of 0.050 to 1.0 millimeters.
Claim 100
Dependent Claim
28 words
An in vitro cell culture method according to claim 7 characterized by providing the solid substrate (A) in the form of a solid body of the calcium compound.
Claim 101
Dependent Claim
26 words
An in vitro cell culture method according to claim 10 characterized in that the solid body is formed of sintered porous granules of the calcium compound.
Claim 102
Dependent Claim
35 words
An in vitro cell culture method according to claim 7, 8, 9, 10 or 11 characterized in that the calcium compound is the hydroxyapatite or tricalcium phosphate form of calcium phosphate or calcium carbonate. DGC1:D
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