Expansion and differentiation of neuronal precursor cells
AssigneeSkin2Neuron Pty Ltd
InventorsMichael VALENZUELA, Tom DUNCAN, An TRUONG
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Abstract
The invention relates to preparation of neuronal precursor cells, compositions comprising same and therapeutic uses.
Claims31
A method for producing a composition of neuronal precursor cells, or of cells capable of proliferation that express neural lineage biomarkers including:
- treating a sample of hair follicle cells in conditions enabling the transition of hair follicle cells to a growth phase, thereby forming a sample of conditioned cells;
- treating the sample of conditioned cells in conditions enabling enrichment of the number of cells containing neuronal lineage biomarkers in the sample;
- thereby producing the composition of neuronal precursor or cells capable of proliferation that express neural lineage biomarkers.
The method of any one of the preceding claims wherein hair follicle cells are provided in a sample of skin.
The method of any one of the preceding claims wherein the hair follicle cells are treated in conditions enabling the majority of the hair follicle precursor cells to transition to a growth phase.
The method of any one of the preceding claims wherein the hair follicle cells are treated with an anti-refractory hair follicle factor for promoting transition of hair follicle cells from telogen to anagen phase, thereby enabling the retention or transition of hair follicle cells in the skin to a growth phase.
The method of any one of the preceding claims wherein the hair follicle cells are treated with a pro-growth factor for promoting transition of hair follicle cells from telogen to anagen phase, thereby enabling the retention or transition of hair follicle cells in the skin to a growth phase.
The method of any one of the preceding claims wherein the hair follicle cells are cultured in cell culture medium for hair follicle cells.
The method of any one of the preceding claims wherein the skin is human skin, or wherein the hair follicles are human hair follicles.
The method of any one of the preceding claims wherein the skin or hair follicles are of the scalp.
The method of any one of the preceding claims wherein the skin or hair follicles are of the midline occipital scalp.
The method of any one of the preceding claims including:
- treating a sample of skin, the skin including hair follicle cells, in conditions enabling the transition of hair follicle cells in the skin to a growth phase, thereby forming a sample of conditioned skin;
- releasing cells from the conditioned skin into the sample;
- depleting terminally differentiated cells or apoptotic cells or cell debris from the sample, thereby forming a sample of non-terminally differentiated cells;
- and optionally
- treating the sample of non-terminally differentiated cells in conditions enabling expansion of the number of non-terminally differentiated cells in the sample;
- thereby producing the composition of neuronal precursor cells or cells capable of proliferation that express neural lineage biomarkers.
The method of any one of the preceding claims wherein at the completion of the depletion step, the sample contains no more than about 5% by number of terminally differentiated cells.
The method of any one of the preceding claims wherein the composition produced by the method has the following characteristics:
- <5% of cells express astroglial GFAP, adiponectin, oligodendrocyte 04, myofibroblast SMA.
- >90% of cells are positive for nestin, CD133 and 13111-tubulin.
The method of any one of the preceding claims wherein the composition produced by the method has a neuronal yield of 90 to 100% following in vitro neuronal differentiation.
The method of any one of the preceding claims wherein the cells of the composition are expandable to produce in the order of 107 homogenous and unipotent neuronal precursor cells within 4 weeks.
Dependencies are inferred from the claim text.