Keratinocytes maintain compartmentalization between dermal papilla and fibroblasts in 3D heterotypic tri‐cultures

Autor: Lifeng Kang, Paul C. Ho, Justin J. Y. Tan, Chunyong Wu, John E.A. Common
Rok vydání: 2019
Předmět:
Zdroj: Cell Proliferation
ISSN: 1365-2184
0960-7722
DOI: 10.1111/cpr.12668
Popis: Objectives Reproducing 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 methods Dermal 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. Results The 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. Conclusions Keratinocytes 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.
Cultures comprising of dermal papilla cells, keratinocytes, and human dermal fibroblasts were prepared from sequential seeding into hydrogel microwells, in order to reproduce the spatial orientation of hair follicular cells in vivo. This work demonstrated the importance of keratinocytes in maintaining cellular compartmentalization and dermal papilla aggregation, which is necessary for hair follicle development and function.
Databáze: OpenAIRE
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