Large-scale cultivation of transplantable dermal papilla cellular aggregates using microfabricated PDMS arrays

In this work we have developed a strategy for cultivating dermal papilla (DP) cells to form multiple arrayed spheroidal microtissues for transplantation on a micropatterned polydimethylsiloxane (PDMS)-based tissue culture polystyrene (TCPS) plate system. We also describe the behavior of dermal papilla cells on this platform and the spontaneous formation of spheroidal microtissues by DP cells. We used a hydrophobic PDMS arrayed chip as a master to separate the seeded cells in the TCPS culture plate. By controlling the cell seeding densities, a microwell with arrayed DP spheroidal microtissues was easily formed. Formation of DP microtissues was associated with overlapping multilayered cells on the microwells and low cell-substrate adhesivity on the PDMS film. The microwell environment enhanced the aggregation of DP cells into spheroidal microtissues on the TCPS culture plate. The spheroidal microtissues preserved their hair induction potential in vitro and in vivo. A large quantity of DP spheroidal microtissues could be obtained rapidly and simply using this platform. We could harvest hundreds of DP microtissues (352 microtissues) with a cell seeding density of 1×10⁶ cells well⁻¹ after 3 days cultivation in one well of a 24-well plate. This is the first demonstration of the formation of DP spheres in large quantitites.