Abstract
ObjectivesReproducing human hair follicles in vitro is often limited by various reasons such as the lack of a systematic approach to culture distinct hair follicle cell types to reproduce their spatial relationship. Here, we reproduce hair follicle‐like constructs resembling the spatial orientation of different cells in vivo, to study the role of keratinocytes in maintaining cellular compartmentalization among hair follicle‐related cells.Materials and methodsDermal papilla (DP) cells, HaCaT keratinocytes and human dermal fibroblast (HDF) cells were seeded sequentially into three‐dimensional (3D) microwells fabricated from polyethylene glycol diacrylate hydrogels. Quantitative polymerase chain reaction was used to compare inductive gene expression of 3D and two‐dimensional (2D) DP. DP and HaCaT cells were transfected with green fluorescent protein and red fluorescent protein lentivirus, respectively, to enable cell visualization using confocal microscopy. ResultsThe 3D DP cultures showed significantly enhanced expression of essential DP genes as compared 2D cultures. Core‐shell configurations containing keratinocytes forming the outer shell and DP forming the core were observed. Migratory polarization was mediated by cell‐cell interaction between the keratinocytes and HDF cells, while preserving the aggregated state of the DP cells.ConclusionsKeratinocytes may play a role in maintaining compartmentalization between the DP and the surrounding HDF residing in the dermis, and therefore maintains the aggregative state of the DP cells, necessary for hair follicle development and function.
Highlights
The hair follicle is often considered a mini organ residing in the der‐ mal layer of the skin
We introduce a systematic approach for producing 3D core‐shell heterotypic spheroids of controlled sizes by the sequential seeding of dissociated human dermal papilla (DP) cells, keratinocytes and human dermal fibroblast (HDF) cells into microarray hydrogels fabricated from poly (ethylene glycol) diacrylate (PEGDA) using soft photolithography
We have further demonstrated, using green fluorescent protein‐expressing DP cells and red fluo‐ rescent protein‐expressing HaCaT keratinocytes to show that the core‐shell structure is achievable using our PEGDA microw‐ ell constructs
Summary
The hair follicle is often considered a mini organ residing in the der‐ mal layer of the skin It is a composite of various specialized and dif‐ ferentiated cells, with each residing in a distinct compartment and having unique functional roles that regulate hair growth, mediated by a series of complex cell‐cell interactions.[1] Research concerning the hair follicles has established several findings essential for un‐ derstanding their biology necessary for recapitulating microenvi‐ ronments suitable for de novo hair follicle neogenesis. It was established that the dermal papilla (DP) cells express greater in vivo like character in their gene and protein expression when cultured in three‐dimensional (3D) microenvironments as compared to flat two‐dimensional (2D) surfaces. We have observed that keratino‐ cytes may have a role in maintaining the aggregative state of the DP cells within the dermis, which could be necessary for the hair follicle development and function
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
