Compositions for the in vitro derivation and culture of embryonic stem (ES) cell lines with germline transmission capability and for the culture of adult stem cells
AssigneeThromboGenics NV
InventorLuc Schoonjans
Life of this patent
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Abstract
The present invention encompasses compositions for deriving, maintaining, and growing pluripotent and germ-line competent mammalian embryonic stem cells, and for deriving, maintaining, and growing adult human stem cells and/or adult early progenitor cells. Such compositions comprising a conditioned medium of a cell line expressing limited amounts of Leukemia Inhibitory Factor (LIF). The media of the present invention are used or for the generation of pluripotent and germ-line competent embryonic stem cells of mammals, and for the generation of adult human stem cells and/or adult early progenitor cells.
Claims11
A method for producing culture medium for the derivation of pluripotent embryonic stem cells from mouse blastocysts, said method comprising the steps of:
- cultivating immortalized rabbit fibroblast cell line Rab9 (ATCC catalogue CRL 1414) in basal embryonic stem cell medium so as to obtain a confluent monolayer culture, and
- collecting from said confluent monolayer culture a stroma-free embryonic stem cell medium conditioned by said immortalized fibroblast cell line, for use in said derivation;
- wherein said conditioned medium contains less than 0.1 ng/ml Leukemia Inhibitory Factor (LIF).
A method for producing culture medium for the derivation of pluripotent embryonic stem cells from primordial germ cells from a postimplantation mouse embryo, said method comprising the steps of:
- cultivating immortalized rabbit fibroblast cell line Rab9 (ATCC catalogue CRL 1414) in basal embryonic stem cell medium so as to obtain a confluent monolayer culture, and
- collecting from said confluent monolayer culture a stroma-free embryonic stem cell medium conditioned by said immortalized fibroblast cell line, for use in said derivation;
- wherein said conditioned medium contains less than 0.1 ng/ml Leukemia Inhibitory Factor (LIF).
A method for producing culture medium for the maintenance of pluripotent embryonic stem cells of mice for at least 10 passages as undifferentiated cells, said method comprising the steps of:
- cultivating immortalized rabbit fibroblast cell line Rab9 (ATCC catalogue CRL 1414) in basal embryonic stem cell medium so as to obtain a confluent monolayer culture, and
- collecting from said confluent monolayer culture a stroma-free embryonic stem cell medium conditioned by said immortalized fibroblast cell line, for use in said maintenance;
- wherein said conditioned medium contains less than 0.1 ng/ml Leukemia Inhibitory Factor (LIF).
A method for producing culture medium for the growth of pluripotent embryonic stem cells of mice for at least 10 passages as undifferentiated cells, said method comprising the steps of:
- cultivating immortalized rabbit fibroblast cell line Rab9 (ATCC catalogue CRL 1414) in basal embryonic stem cell medium so as to obtain a confluent monolayer culture, and
- collecting from said confluent monolayer culture a stroma-free embryonic stem cell medium conditioned by said immortalized fibroblast cell line, for use in said growth;
- wherein said conditioned medium contains less than 0.1 ng/ml Leukemia Inhibitory Factor (LIF).
A method for producing culture medium for the expansion of human umbilical cord blood (UCB) CD34+ cells, said method comprising the steps of:
- cultivating immortalized rabbit fibroblast cell line Rab9 (ATCC catalogue CRL 1414) in basal embryonic stem cell medium so as to obtain a confluent monolayer culture, and
- collecting from said confluent monolayer culture a stroma-free embryonic stem cell medium conditioned by said immortalized fibroblast cell line, for use in said expansion;
- wherein said conditioned medium contains less than 0.1 ng/ml Leukemia Inhibitory Factor (LIF).
Dependencies are inferred from the claim text.