title: Raman spectroscopy: a noninvasive analysis tool for the discrimination of human skin cells.
abstract: Noninvasive monitoring of tissue-engineered (TE) constructs during their in vitro maturation or postimplantation in vivo is highly relevant for graft evaluation. However, traditional methods for studying cell and matrix components in engineered tissues such as histology, immunohistochemistry, or biochemistry require invasive tissue processing, resulting in the need to sacrifice of TE constructs. Raman spectroscopy offers the unique possibility to analyze living cells label-free in situ and in vivo solely based on their phenotype-specific biochemical fingerprint. In this study, we aimed to determine the applicability of Raman spectroscopy for the noninvasive identification and spectral separation of primary human skin fibroblasts, keratinocytes, and melanocytes, as well as immortalized keratinocytes (HaCaT cells). Multivariate analysis of cell-type-specific Raman spectra enabled the discrimination between living primary and immortalized keratinocytes. We further noninvasively distinguished between fibroblasts, keratinocytes, and melanocytes. Our findings are especially relevant for the engineering of in vitro skin models and for the production of artificial skin, where both the biopsy and the transplant consist of several cell types. To realize a reproducible quality of TE skin, the determination of the purity of the cell populations as well as the detection of potential molecular changes are important. We conclude therefore that Raman spectroscopy is a suitable tool for the noninvasive in situ quality control of cells used in skin tissue engineering applications.
authors.affiliation: Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Stuttgart, Germany.
authors.forename: Marieke
authors.initials: M
authors.lastname: Pudlas
authors.forename: Steffen
authors.initials: S
authors.lastname: Koch
authors.forename: Carsten
authors.initials: C
authors.lastname: Bolwien
authors.forename: Sibylle
authors.initials: S
authors.lastname: Thude
authors.forename: Nele
authors.initials: N
authors.lastname: Jenne
authors.forename: Thomas
authors.initials: T
authors.lastname: Hirth
authors.forename: Heike
authors.initials: H
authors.lastname: Walles
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
mesh_terms: D000328:Adult; D002448:Cell Adhesion; D020405:Dermis / Q000166:cytology; D000078404:Epidermal Cells; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D066298:In Vitro Techniques; D015603:Keratinocytes / Q000166:cytology; D008297:Male; D008544:Melanocytes / Q000166:cytology; D010641:Phenotype; D025341:Principal Component Analysis; D012867:Skin / Q000166:cytology*; D013059:Spectrum Analysis, Raman / Q000379:methods*