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Controllable production of transplantable adult human high-passage dermal papilla spheroids using 3D Matrigel culture
We have succeeded in culturing human dermal papilla (DP) cells spheroids and developed a three-dimensional Matrigel (basement membrane matrix) culture technique that can enhance and restores DP cells unique characteristics in vitro.
When 10000 DP cells were cultured on the 96 well plates pre-coated with Matrigel for 5 days, both passage 2 and passage 8 DP cells formed spheroidal microtissues with a diameter of 150-250 μm in an aggregative and proliferative manner. We transferred and re-cultured these DP spheroids onto commercial plates. Cells within DP spheres could disaggregate and migrate out, which was similar to primary DP. Moreover, we examined the expression of several genes and proteins associated with hair follicle inductivity of DP cells, such as NCAM, Versican and α-SMA, and confirmed that their expression level was elevated in the spheres compared with the dissociated DP cells. To examine hair-inducing ability of DP spheres, hair germinal matrix cells and DP spheres were mixed and cultured on Matrigel. Unlike the dissociated DP cells and hair germinal matrix cells co-cultured in two dimensions, hair germinal matrix cells can differentiate into hair-like fibers under the induction of the DP spheres made from the high passage cells (passage 8) in vitro.
We are the first to show that passage 3 human hair germinal matrix cells differentiate into hair-like fiber in the presence of human DP spheroids.
These results suggest that three-dimensional Matrigel culture technique is an ideal culture model for forming DP spheroids and that sphere formation partially models the intact DP, resulting in hair induction, even by high passage DP cells.
We have succeeded in culturing human dermal papilla (DP) cells spheroids and developed a three-dimensional Matrigel (basement membrane matrix) culture technique that can enhance and restores DP cells unique characteristics in vitro.
When 10000 DP cells were cultured on the 96 well plates pre-coated with Matrigel for 5 days, both passage 2 and passage 8 DP cells formed spheroidal microtissues with a diameter of 150-250 μm in an aggregative and proliferative manner. We transferred and re-cultured these DP spheroids onto commercial plates. Cells within DP spheres could disaggregate and migrate out, which was similar to primary DP. Moreover, we examined the expression of several genes and proteins associated with hair follicle inductivity of DP cells, such as NCAM, Versican and α-SMA, and confirmed that their expression level was elevated in the spheres compared with the dissociated DP cells. To examine hair-inducing ability of DP spheres, hair germinal matrix cells and DP spheres were mixed and cultured on Matrigel. Unlike the dissociated DP cells and hair germinal matrix cells co-cultured in two dimensions, hair germinal matrix cells can differentiate into hair-like fibers under the induction of the DP spheres made from the high passage cells (passage 8) in vitro.
We are the first to show that passage 3 human hair germinal matrix cells differentiate into hair-like fiber in the presence of human DP spheroids.
These results suggest that three-dimensional Matrigel culture technique is an ideal culture model for forming DP spheroids and that sphere formation partially models the intact DP, resulting in hair induction, even by high passage DP cells.
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