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US 2024/0392318 A1Published application
Host cell lines and methods for identifying and using such host cell lines
AssigneeSanofi
InventorsBarak Balva, Christine DeMaria, Karen Vincent, Saurabh Sen, Patricia Alves, Mafalda Dias, Jose Escandell, Daniel Pais
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Abstract
Disclosed are host cell lines and producer cell lines derived from a HeLaS3 parental cell lines and methods of producing the host cell lines and producer cell lines.
Claims30
A host cell derived from a HeLaS3 parental cell line.
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A method for selecting a host cell line, comprising:
- (a) expanding one or more population(s) of a HeLaS3 parental cell line in a serum-free media;
- (b) selecting and isolating one or more single cell clones from step (a);
- (c) expanding each of the one or more single cell clones from step (b) in a serum-free media;
- (d) selecting from each of the one or more single cell clones from step (c) and analyzing the one or more single cell clones for at least one of the following characteristics:
- (i) cell doubling time,
- (ii) transfection efficiency,
- (iii) peak viable cell density;
- (iv) cell outgrowth after seeding at low cell density;
- (v) cell viability:
- (vi) clumping degree;
- or
- (vii) any combination thereof;
- and
- (e) growing the one or more single cell clones selected from step (d) in a serum-free media.
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A method of producing a host cell line comprising the steps of:
- (a) expanding one or more population(s) of a HeLaS3 parental cell line at a density of 0.5 cells/well in media comprising EX-CELL HeLa growth medium supplemented with 6 mM L-Glutamine, 50% DMEM/F-12 (with 6 mM L-Glutamine), 20% conditioned media and 1× InstiGRO CHO supplement, wherein all components of the media and conditioned media are serum free and free of animal derived components;
- (b) selecting and isolating one or more single cell clones from step (a);
- (c) expanding each of the one or more single cell clones from step (b) in a serum-free media;
- (d) selecting from each of the one or more single cell clones from step (c) for at least one of the following characteristics:
- (i) cell viability,
- (ii) cell doubling time,
- (iii) transfection efficiency,
- (iv) peak viable cell density;
- (v) clumping degree;
- (vi) cell outgrowth after seeding at low cell density;
- and
- (vii) any combination thereof;
- and
- (e) isolating and expanding the one or more single cell clones selected from step (d) in a serum-free media thereby producing the host cell line.
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Dependencies are inferred from the claim text.