Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Primate cell – per se
Patent
1998-04-28
1999-12-28
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Primate cell, per se
435 11, 435370, 435378, 435383, 435391, 435392, 435397, 424 937, C12N 500, A01N 102
Patent
active
060080477
ABSTRACT:
The present invention provides a method for producing an expanded non-transformed cell culture of human liver cells comprising the steps of: (1) preparing partially purified, minced human liver tissue, (2) concentrating the resulting cells and tissue pieces, (3) resuspending the concentrated tissue cells and pieces in a growth medium, (4) culturing the resuspended cells in the growth medium for a time and under conditions to effect sustained cell division, and (5) passaging the cultured human liver cells periodically to expand the culture. The growth medium comprises a combination of a basal medium and ingredients to provide a medium in which the cultured human liver cells are selectively proliferated without being transformed, providing an expanded culture of proliferated, functionally differentiated human liver cells that is substantially free of fibroblast, macrophage and capillary endothelial cells. Also provided is the improvement of harvesting cells of the expanded culture at a selected PDL preferably>5, providing a high density cell suspension of such proliferated human liver cells, and incubating such high density cell suspension in a calm-down medium to induce a mitotically quiescent state and, using a culture procedure which encourages aggregation, making the cells adhere tightly to form a three-dimensional cell organization typical of the organ of origin, thereby forming organoids.
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Ambesi-Impiombato F. Saverio
Coon Hayden G.
Curcio Francesco
Bundock Jr. John P.
Lankford , Jr. Leon B.
Livercell L.L.C.
Tate Christopher R.
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