Methods for manufacture of neuronal precursors

A method for determining whether source tissue is suitable for providing HFNs, the method comprising:

• providing a reference glucose consumption profile from intact human hair follicles that have been demonstrated to generate a viable HFN cell line;
• providing source tissue in the form of a sample of hair follicle cells;
• determining the glucose consumption of the follicles in the sample;
• comparing the glucose consumption of the follicles in the sample to the reference glucose consumption profile;
• determining that the source tissue is likely not suitable for providing HFNs if the glucose consumption of the follicles is the lower than that of the reference glucose consumption profile.

A method for determining the likelihood that a sample of hair follicles comprises sufficient numbers of HFNs for isolation, the method comprising:

• providing a sample of hair follicles;
• determining the glucose consumption of the follicles in the sample;
• comparing the glucose consumption of the follicles in the sample to a reference glucose consumption profile from intact human hair follicles from which HFNs were successfully isolated;
• determining that the sample of hair follicles likely comprises sufficient numbers of HFNs for isolation, if the glucose consumption of the follicles in the sample is the same or higher than that of the reference glucose consumption profile.

A method for determining the likelihood that a sample of hair follicles comprises sufficient numbers of HFNs for isolation, the method comprising:

• providing a sample of hair follicles;
• determining the glucose consumption of the follicles in the sample;
• comparing the glucose consumption of the follicles in the sample to a reference glucose consumption profile from intact human hair follicles from which HFNs were successfully isolated;
• determining that the sample of hair follicles likely does not comprise sufficient numbers of HFNs for isolation, if the glucose consumption of the follicles is the lower than that of the reference glucose consumption profile.

An improved method for the generation of neuronal precursor cells from a sample of hair follicles, the method comprising:

• 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 follicles;
• determining the glucose consumption of the conditioned follicles;
• comparing the glucose consumption of the conditioned follicles to a reference glucose consumption profile representative of hair follicles from which viable HFNs were isolated;
• proceeding to treat the conditioned follicles in conditions enabling enrichment of the number of cells expressing neuronal lineage biomarkers, if the glucose consumption of the conditioned follicles is the same or higher than that of the reference glucose consumption profile,
• thereby generating a composition of neuronal precursor cells from a sample of hair follicles.

An improved method for the generation of neuronal precursor cells from a sample of hair follicle cells, the method comprising:

• treating a sample of hair follicle cells in conditions enabling the transition of hair follicles to a growth phase, thereby forming a sample of conditioned follicles;
• determining the glucose consumption of the conditioned follicles;
• comparing the glucose consumption of the conditioned follicles to a reference glucose consumption profile representative of hair follicles from which viable HFNs were not successfully isolated;
• proceeding to treat the conditioned follicles in conditions enabling enrichment of the number of cells containing neuronal lineage biomarkers, if the glucose consumption of the conditioned follicles is higher than that of the reference glucose consumption profile,
• thereby generating a composition of neuronal precursor cells from a sample of hair follicles.