FIG 1. de novo Organoids in a 24 well plate

Diagnostics/Assays only offered to physicians after regulatory clearance.

Alopecia is often diagnosed at a rather late stage of the disease progression. We aim at further developing our hair  organoid technology to be able to predict disease susceptibility long before the first symptoms become observable. This approach includes:

Personalized - INVITRO- drug testing

For any indication where more than one drug is available, pre-testing on our personalized INVITRO application (and on the planned personalized hair-on-a-chip devices) is conceivable, so that only the drug which showed maximal effect is then prioritized for the patient.

Available assays:

1) Oxidative stress with fluorescent probe.

To evaluate the general levels of oxidative stress species (ROS), a fluorescent dye-based probe will be used.

Follicles/ hair samples stored in an sterile vial with 5ml Thermosol will be treated with the probe and imaged with high-throughput confocal microscope (Operetta, PerkinElmer). A minimum of 3 follicles/ hair samples will be used for the assessment. 3 follicles / hair samples be required for the positive control.

Assay analysis: imaged-based, average fluorescent values in positive cells normalized on the values of follicles treated with a positive control (stress inducer). This normalization will indicate the sample status in relation to the maximum oxidative levels. This will be substitute/complemented it with average values from healthy subjects when available. The assay is based on INVTROHAIR proprietary algorithms and htp-microscopy.

Total needed follicles/ hair samples minimum 6.

Time needed: 3 weeks

2) Viability assay.

To evaluate the status of the cells, we will perform a viability assay. This will be tested with 2 kits of an INVITROHAIR proprietary cell Viability Assay.

Follicles/ hair samples store in Thermosol will be treated according to the manufacturer’s instructions.

Assay analysis: absorbance/luminescence-based, average values will be related to the values of follicles treated with a positive control (cell death inducer). This normalization will indicate the sample status in relation to the maximum cell death status. This will be substituted/complemented it with average values from healthy subjects when available.

Total needed follicles/ hair samples minimum 6.

Time needed: 3 weeks

3) Proliferative activity of the stem cell pool -ONLY WITH HAIR FOLLICLES-.

To evaluate the proliferative status of the stem cells in the hair follicles, the following staining will be performed:

Ki67; Hoechs; SOX2 and SOX9. Casp3

Total needed hair follicles minimum 8

Time needed: 3 weeks

FIG.2 refers to the conduction of the Stemness Assay as a part of our P4 Program.