Abstract
We developed a human skin equivalent (HSE) containing blood and lymph-like capillary networks using a cell-coating technique. This technique is a rapid fabrication technology of three-dimensional cellular constructs by cell surface coating using layer-by-layer assembled nanofilms of extracellular matrices. The thickness of a dermis consisting of normal human dermal fibroblast was easily controlled from approximately 5–100 μm by altering the seeded cell number. Keratinocytes as an epidermis showed homogeneous differentiation on the surface of the dermis by lifting to air-liquid interface, and immunological staining represented distinct four layers, stratum basale, spinosum, granulosum, and corneum. Interestingly, the measurement of transepithelial electrical resistance (TEER) indicated prolongation of the daily reached maximum value of TEER with an increase in the numbers of dermis layers. HSEs with six layers of dermis revealed the longest period maintained, over 500 Ω cm2 of TEER. The co-sandwich culture of human umbilical vein endothelial cells and normal human dermal lymphatic microvascular endothelial cells within an eight-layered dermis showed in vitro co-network formation of individual blood and lymph-like capillaries inside the dermis. This is the first report on homogeneous full-thickness HSEs with blood and lymph capillary networks that will be useful for biomedical and pharmaceutical applications.
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Acknowledgments
This work was supported mainly by PRESTO-JST, and partly by a Grant-in-Aid for Scientific Research on Innovative Areas (21106514 and 26106717) from MEXT of Japan, The Noguchi Institute Fund, and by NEXT Program from JSPS (LR026).
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Matsusaki, M., Fujimoto, K., Shirakata, Y., Hirakawa, S., Hashimoto, K., Akashi, M. (2017). Development of Full-Thickness Human Skin Equivalents with Blood and Lymph-like Capillary Networks by Cell Coating Technology. In: Sugibayashi, K. (eds) Skin Permeation and Disposition of Therapeutic and Cosmeceutical Compounds. Springer, Tokyo. https://doi.org/10.1007/978-4-431-56526-0_31
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DOI: https://doi.org/10.1007/978-4-431-56526-0_31
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