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US 8,133,727 B2Granted
In vitro human B lymphopoiesis culture system
AssigneeCalifornia Institute of Technology
InventorsXin Luo, Lili Yang, David Baltimore
Life of this patent
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Abstract
The present disclosure relates to systems, methods and compositions for the generation of antibody-producing B cells in vitro. Some embodiments are related to an in vitro system for generating antibody-producing B cells from hematopoietic stem/progenitor cells (HSPCs).
Claims22
A method for generating a population of antibody-producing B cells in vitro, comprising:
- (a) contacting a population of hematopoietic stem/progenitor cells (HSPCs) in vitro with a polynucleotide delivery system, wherein the polynucleotide delivery system comprises a polynucleotide encoding an antibody of interest;
- (b) culturing the HSPCs in the presence of one or more B-cell priming factors;
- (c) co-culturing the HSPCs with a population of first supporting cells expressing one or more B-lineage growth factors until at least about 20% of the HSPCs become CD19+μ+;
- (d) co-culturing the CD19+μ+ cells obtained in step (c) with a population of second supporting cells expressing CD40L in the presence of one or more B-cell activators until at least about 20% of the CD19+μ+ cells become B cells that produce the antibody of interest.
1 more element
A method for generating a population of antibody-producing B cells in vitro, comprising:
- (a) culturing a population of hematopoietic stem/progenitor cells (HSPCs) in the presence of one or more B-cell priming factors;
- (b) co-culturing the HSPCs with a population of first supporting cells expressing one or more B-lineage growth factors until at least about 20% of the HSPCs become CD19+μ+;
- (c) co-culturing the CD19+μ+ cells obtained in step (b) with a population of second supporting cells expressing CD40L in the presence of one or more B-cell activators until at least about 20% of the CD19+μ+ cells become antibody-producing B cells.
A method for generating antibody-producing B cells, comprising:
- transfecting a population of target cells in vitro with a polynucleotide delivery system, where the polynucleotide delivery system comprises a polynucleotide encoding an antibody of interest;
- co-culturing the target cells with a population of supporting cells expressing CD40L in the presence of one or more B-cell activators until at least about 20% of the target cells become B cells that produce the antibody of interest.
Dependencies are inferred from the claim text.