Methods of generating brain organoids and repairing non-viable embryoid bodies
AssigneeUniversity of Saskatchewan
InventorsTyler Joseph Richard WENZEL, Darrell MOUSSEAU
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Abstract
The disclosure relates to methods of generating at least one brain organoid-sufficient embryoid body or repairing at least one non-viable embryoid body, the method comprising incubating a population of induced pluripotent stem cells (iPSCs) in an embryoid body formation medium (EB FM), the EB FM comprising a culture medium, glutamine, recombinant human insulin, recombinant human transferrin, sodium selenite, and thermostable fibroblast growth factor 2 (FGF2). The disclosure further relates to methods of generating brain organoids from brain organoid-sufficient embryoid bodies and repairing embryoid bodies.
Claims32
A method of generating at least one brain organoid-sufficient embryoid body, the method comprising incubating a population of induced pluripotent stem cells (iPSCs) in an embryoid body formation medium (EB FM), the EB FM comprising:
- a cell culture medium, optionally Dulbecco's modified Eagle medium (DMEM)
- about 1 μM to about 100 mM glutamine, optionally about 0.1 mM to about 10 mM L-alanyl-L-glutamine dipeptide;
- about 5 μg/ml to about 30 μg/ml of recombinant human insulin;
- about 5 μg/ml to about 30 μg/ml of recombinant human transferrin;
- about 5 ng/ml to about 30 ng/ml of sodium selenite;
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- about 1 ng/ml to about 200 ng/ml, optionally about 10 ng/ml to about 25 ng/ml, of thermostable fibroblast growth factor 2 (FGF2).
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A method of repairing at least one non-viable embryoid body, the method comprising:
- obtaining a population of embryoid bodies (EBs) comprising at least one non-viable embryoid body, and
- incubating the population of embryoid bodies, optionally for at least about 24 hours, with an embryoid body formation medium (EB FM) comprising:
- a cell culture medium, optionally Dulbecco's modified Eagle medium (DMEM);
- about 1 μM to about 100 mM glutamine, optionally about 0.1 mM to about 10 mM L-alanyl-L-glutamine dipeptide;
- about 5 μg/ml to about 30 μg/ml of recombinant human insulin;
- about 5 μg/ml to about 30 μg/ml of recombinant human transferrin;
- and
- about 5 ng/ml to about 30 ng/ml of sodium selenite.
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Dependencies are inferred from the claim text.