Organotypic 3D Skin Models: Human Epidermal Equivalent Cultures from Primary Keratinocytes and Immortalized Keratinocyte Cell Lines

2020 ◽  
pp. 45-61
Author(s):  
Gijs Rikken ◽  
Hanna Niehues ◽  
Ellen H. van den Bogaard
Keyword(s):  
Cell Lines ◽  
Primary Keratinocytes ◽  
Keratinocyte Cell ◽  
3D Skin

scholarly journals Effects of sunitinib on the expression of keratin 9 in human keratinocyte cell lines and a 3D skin model

2018 ◽  
Vol WCP2018 (0) ◽  
pp. PO4-9-30
Author(s):  
Ayaka Yoshida ◽  
Kazuhiro Yamamoto ◽  
Akane Murakawa ◽  
Takahiro Ishida ◽  
Tsutomu Nakagawa ◽  
...  
Keyword(s):  
Cell Lines ◽  
Skin Model ◽  
Human Keratinocyte ◽  
Keratinocyte Cell ◽  
3D Skin

scholarly journals hTERT-Driven Immortalization of RDEB Fibroblast and Keratinocyte Cell Lines Followed by Cre-Mediated Transgene Elimination

2021 ◽  
Vol 22 (8) ◽  
pp. 3809
Author(s):  
Nadezhda A. Evtushenko ◽  
Arkadii K. Beilin ◽  
Erdem B. Dashinimaev ◽  
Rustam H. Ziganshin ◽  
Anastasiya V. Kosykh ◽  
...  
Keyword(s):  
Cell Lines ◽  
Ectopic Expression ◽  
Protein Patterns ◽  
Skin Equivalents ◽  
Type Vii Collagen ◽  
Recessive Form ◽  
Immortalized Cells ◽  
Keratinocyte Cell ◽  
Cell Changes ◽  
3D Skin

The recessive form of dystrophic epidermolysis bullosa (RDEB) is a crippling disease caused by impairments in the junctions of the dermis and the basement membrane of the epidermis. Using ectopic expression of hTERT/hTERT + BMI-1 in primary cells, we developed expansible cultures of RDEB fibroblasts and keratinocytes. We showed that they display the properties of their founders, including morphology, contraction ability and expression of the respective specific markers including reduced secretion of type VII collagen (C7). The immortalized keratinocytes retained normal stratification in 3D skin equivalents. The comparison of secreted protein patterns from immortalized RDEB and healthy keratinocytes revealed the differences in the contents of the extracellular matrix that were earlier observed specifically for RDEB. We demonstrated the possibility to reverse the genotype of immortalized cells to the state closer to the progenitors by the Cre-dependent hTERT switch off. Increased β-galactosidase activity and reduced proliferation of fibroblasts were shown after splitting out of transgenes. We anticipate our cell lines to be tractable models for studying RDEB from the level of single-cell changes to the evaluation of 3D skin equivalents. Our approach permits the creation of standardized and expandable models of RDEB that can be compared with the models based on primary cell cultures.

scholarly journals Primary keratinocytes have an adhesion dependent S phase checkpoint that is absent in immortalized cell lines

Oncogene ◽  
1998 ◽  
Vol 17 (24) ◽  
pp. 3083-3092 ◽  
Author(s):  
Paul J Hauser ◽  
Deepak Agrawal ◽  
W J Pledger
Keyword(s):  
Cell Lines ◽  
S Phase ◽  
Primary Keratinocytes ◽  
Immortalized Cell Lines ◽  
Immortalized Cell ◽  
S Phase Checkpoint

scholarly journals Characterization of human keratinocyte cell lines for barrier studies

2021 ◽  
pp. 100018
Author(s):  
Mary C. Moran ◽  
Radha P. Pandya ◽  
Kimberly A. Leffler ◽  
Takeshi Yoshida ◽  
Lisa A. Beck ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Keratinocyte ◽  
Keratinocyte Cell

scholarly journals A role for the E-cadherin cell-cell adhesion molecule during tumor progression of mouse epidermal carcinogenesis.

1991 ◽  
Vol 115 (2) ◽  
pp. 517-533 ◽  
Author(s):  
P Navarro ◽  
M Gómez ◽  
A Pizarro ◽  
C Gamallo ◽  
M Quintanilla ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Lines ◽  
Spindle Cell ◽  
Inverse Correlation ◽  
Malignant Cell ◽  
Mouse Tumors ◽  
Keratinocyte Cell ◽  
Well Differentiated ◽  
E Cadherin ◽  
Cell Cell

The expression of the cell-cell adhesion molecules E- and P-cadherin has been analyzed in seven mouse epidermal keratinocyte cell lines representative of different stages of epidermal carcinogenesis. An inverse correlation between the amount of E-cadherin protein and tumorigenicity of the cell lines has been found, together with a complete absence of E-cadherin protein and mRNA expression in three carcinoma cell lines (the epithelioid HaCa4 and the fibroblastoid CarB and CarC cells). A similar result has been detected in tumors induced in nude mice by the cell lines, where induction of E-cadherin expression takes place in moderately differentiated squamous cell carcinomas induced by HaCa4 cells, although at much lower levels than in well-differentiated tumors induced by the epithelial PDV or PDVC57 cell lines. Complete absence of E-cadherin expression has been observed in spindle cell carcinomas induced by CarB or CarC cells. P-cadherin protein was detected in all cell lines that exhibit an epithelial (MCA3D, AT5, PDV, and PDVC57) or epithelioid (HaCa4) morphology, as well as in nude mouse tumors, independent of their tumorigenic capabilities. However, complete absence of P-cadherin was observed in the fibroblast-like cells (CarB and CarC) and in spindle cell carcinomas. The introduction of an exogenous E-cadherin cDNA into HaCa4 cells, or reactivation of the endogenous E-cadherin gene, leads to a partial suppression of the tumorigenicity of this highly malignant cell line. These results suggest a role for E-cadherin in the progression to malignancy of mouse epidermal carcinogenesis. They also suggest that the loss of both E- and P-cadherin could be associated to the final stage of carcinogenesis, the development of spindle cell carcinomas.

scholarly journals The Selection of NFκB Inhibitors to Block Inflammation and Induce Sensitisation to FasL-Induced Apoptosis in HNSCC Cell Lines Is Critical for Their Use as a Prospective Cancer Therapy

2019 ◽  
Vol 20 (6) ◽  
pp. 1306 ◽  
Author(s):  
Mario Scheurer ◽  
Roman Brands ◽  
Mohamed El-Mesery ◽  
Stefan Hartmann ◽  
Urs Müller-Richter ◽  
...  
Keyword(s):  
Cancer Therapy ◽  
Cell Lines ◽  
In Vitro Study ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell ◽  
Nfκb Pathway ◽  
Induced Apoptosis ◽  
Selection Of ◽  
Hnscc Cell

Inflammation is a central aspect of tumour biology and can contribute significantly to both the origination and progression of tumours. The NFκB pathway is one of the most important signal transduction pathways in inflammation and is, therefore, an excellent target for cancer therapy. In this work, we examined the influence of four NFκB inhibitors—Cortisol, MLN4924, QNZ and TPCA1—on proliferation, inflammation and sensitisation to apoptosis mediated by the death ligand FasL in the HNSCC cell lines PCI1, PCI9, PCI13, PCI52 and SCC25 and in the human dermal keratinocyte cell line HaCaT. We found that the selection of the inhibitor is critical to ensure that cells do not respond by inducing counteracting activities in the context of cancer therapy, e.g., the extreme IL-8 induction mediated by MLN4924 or FasL resistance mediated by Cortisol. However, TPCA1 was qualified by this in vitro study as an excellent therapeutic mediator in HNSCC by four positive qualities: (1) proliferation was inhibited at low μM-range concentrations; (2) TNFα-induced IL-8 secretion was blocked; (3) HNSCC cells were sensitized to TNFα-induced cell death; and (4) FasL-mediated apoptosis was not disrupted.

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