Method Profile
Spinal-cord-patterned human spheroids allow sequential measurement of myelin damage, debris clearance, new oligodendrocytes and repair-associated myelination.
Study defined phases of demyelination and repair, and test whether a perturbation changes new-cell myelination in a human-cell model.
Human iPSC-derived spinal-cord-patterned spheroids from 18a or C005 lines, primarily 18a; separately differentiated macrophage-like precursors supplied the microglia-like population.
Long-term spheroid culture with LPC injury and follow-up at four, eight and 12 weeks; clemastine fumarate was tested at 1 μM for eight weeks.
Spheroid and injury model
Human neural spheroids with oligodendrocytes, neurons, astrocytes and introduced microglia-like cells; LPC compared with vehicle.
Repair measurements
EdU labeling, cell and myelin markers, transmission electron microscopy and focused-ion-beam electron microscopy.
Drug comparison
Clemastine fumarate at 1 μM, compared with basal medium and growth-factor-supported conditions.
Confirm damage and debris clearance, then combine EdU, cellular and myelin markers with electron microscopy. For the drug comparison, retain the reported denominator of newly generated oligodendrocytes.
Follow LPC-induced injury and repair for up to 12 weeks; compare clemastine with basal and growth-factor conditions over eight weeks.
Key recovery analyses used five to six spheroids per condition across two differentiations. The clemastine comparison used four to seven spheroids per condition across two differentiations.
LPC versus vehicle; post-injury time points; basal medium, growth-factor-supported conditions and clemastine, with uninjured controls.
New oligodendrocytes and compact but thinner myelin appeared after injury. Clemastine increased the proportion of new oligodendrocytes that myelinated under the tested conditions.
A spinal-cord-patterned human neural spheroid model followed myelin damage, debris clearance and new myelin formation over weeks. In a small drug comparison, clemastine increased the share of newly formed oligodendrocytes that became myelinating cells, a cellular result rather than evidence of clinical efficacy.
