An In Vitro Model of Differentiated Human Airway Epithelia: Methods for Establishing Primary Cultures

2003 ◽  
pp. 115-137 ◽  
Author(s):  
Philip H. Karp ◽  
Thomas O. Moninger ◽  
S. Pary Weber ◽  
Tamara S. Nesselhauf ◽  
Janice L. Launspach ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Primary Cultures ◽  
Airway Epithelia ◽  
Human Airway ◽  
Vitro Model ◽  
Human Airway Epithelia

scholarly journals An in vitro feasibility investigation considering primary human melanocytes for spray- grafting of freshly isolated autologous skin cells for burn treatment and a clinical case report

2019 ◽  
pp. 1-8
Author(s):  
Jörg C. Gerlach ◽  
C. Johnen ◽  
B. Hartmann ◽  
J. Plettig ◽  
K. Bräutigam ◽  
...  
Keyword(s):  
Cell Morphology ◽  
Skin Color ◽  
In Vitro Model ◽  
Primary Cultures ◽  
Cell Suspensions ◽  
Epidermal Keratinocytes ◽  
Cell Counts ◽  
Skin Coloration ◽  
Vitro Model

A skin cell-spray grafting technique that enables the on-site application of freshly isolated autologous single cell suspensions was already applied in many cases on caucasian patients with low skin coloration. Our project hypothesis is that these suspensions contain keratinocytes and vital melanocytes, that are of particular interest for the treatment of patients of darker skin color. To test this, we applied an in vitro model, wherein the feasibility of i) isolating and ii) spraying of freshly isolated autologous melanocyte-keratinocyte cell suspensions was investigated. Primary human epidermal keratinocytes (HEKs) and melanocytes (MCs) were isolated from skin biopsies (n=8). Biochemical parameter, cell counts, cell morphology, growth behavior and immunofluorescence results were compared in two groups using MC cultures and co-cultures of MCs with HEKs. Case information on using the method clinically with one patient is included. The sprayed mixed cell suspensions proliferated in all groups without measurable loss of viability, and MCs exhibited a regular cell morphology in monoculture up to passage 4°. The sprayed MCs and HEKs demonstrated in vitro glucose and lactate metabolism that was comparable to the pipetted controls. In co-culture, well distributed CK14+ HEKs and NKI/beteb+ MCs could be demonstrated, which interacted in the in vitro model. The ratio of HEKs : MCs in our primary cultures were microscopically counted (n=8 each) as mean +/- SD 1,211,000 (+/- 574,343) HEK : 99,625 (+/- 59,025) MC; i.e., a ratio of approx. 12 : 1. Using the isolation method clinically for a patient with dark skin coloration after suffering severe second-degree burns shows a satisfying re-pigmentation of the resulting wound post healing. Freshly isolated spray-on melanocyte/keratinocyte suspensions provide for a considerable amount of viable HEKs and MCs. Using MCs in spray-grafting suspensions could represent a promising approach for treating severe partial-thickness burns and innovative therapy developments that also aim to address cosmetic aspects.

scholarly journals Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0181058 ◽  
Author(s):  
Patricia A. Noguera ◽  
Bianka Grunow ◽  
Matthias Klinger ◽  
Katherine Lester ◽  
Bertrand Collet ◽  
...  
Keyword(s):  
Atlantic Salmon ◽  
In Vitro Model ◽  
Primary Cultures ◽  
Host Pathogen Interactions ◽  
Host Pathogen ◽  
Vitro Model

scholarly journals Endotoxin responsiveness of human airway epithelia is limited by low expression of MD-2

2004 ◽  
Vol 287 (2) ◽  
pp. L428-L437 ◽  
Author(s):  
Hong Peng Jia ◽  
Joel N. Kline ◽  
Andrea Penisten ◽  
Michael A. Apicella ◽  
Theresa L. Gioannini ◽  
...  
Keyword(s):  
Primary Cultures ◽  
Airway Epithelial Cells ◽  
Response To Treatment ◽  
Airway Epithelial ◽  
Airway Epithelia ◽  
Human Airway ◽  
Tnf Α ◽  
Pulmonary Macrophages ◽  
Well Differentiated ◽  
Human Airway Epithelia

The expression of inducible antimicrobial peptides, such as human β-defensin-2 (HBD-2) by epithelia, comprises a component of innate pulmonary defenses. We hypothesized that HBD-2 induction in airway epithelia is linked to pattern recognition receptors such as the Toll-like receptors (TLRs). We found that primary cultures of well-differentiated human airway epithelia express the mRNA for TLR-4, but little or no MD-2 mRNA, and display little HBD-2 expression in response to treatment with purified endotoxin ± LPS binding protein (LBP) and soluble CD14. Expression of endogenous MD-2 by transduction of airway epithelial cells with an adenoviral vector encoding MD-2 or extracellular addition of recombinant MD-2 both increased the responses of airway epithelia to endotoxin + LBP and sCD14 by >100-fold, as measured by NF-κB-luciferase activity and HBD-2 mRNA expression. MD-2 mRNA could be induced in airway epithelia by exposure of these cells to specific bacterial or host products (e.g., killed Haemophilus influenzae, the P6 outer membrane protein from H. influenzae, or TNF-α + IFN-γ). These findings suggest that MD-2, either coexpressed with TLR-4 or secreted when produced in excess of TLR-4 from neighboring cells, is required for airway epithelia to respond sensitively to endotoxin. The regulation of MD-2 expression in airway epithelia and pulmonary macrophages may serve as a means to modify endotoxin responsiveness in the airway.

An in vitro model of human airway epithelium (EpiAirway) for in vitro metabolism and toxicity screening

2007 ◽  
Vol 172 ◽  
pp. S79 ◽  
Author(s):  
Patrick Hayden ◽  
Joseph Kubilus ◽  
Helena Kandárová ◽  
Mitchell Klausner ◽  
George Jackson ◽  
...  
Keyword(s):  
Airway Epithelium ◽  
In Vitro Model ◽  
In Vitro Metabolism ◽  
Toxicity Screening ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
Vitro Model

KGF alters gene expression in human airway epithelia: potential regulation of the inflammatory response

2001 ◽  
Vol 6 (2) ◽  
pp. 81-89 ◽  
Author(s):  
LAWRENCE S. PRINCE ◽  
PHILIP H. KARP ◽  
THOMAS O. MONINGER ◽  
MICHAEL J. WELSH
Keyword(s):  
Keratinocyte Growth Factor ◽  
Primary Cultures ◽  
Airway Epithelia ◽  
Altered Expression ◽  
Human Airway ◽  
Gene Transcripts ◽  
Induced Genes ◽  
Well Differentiated ◽  
Cf Transmembrane Conductance Regulator ◽  
Human Airway Epithelia

Keratinocyte growth factor (KGF) regulates several functions in adult and developing lung epithelia; it causes proliferation, stimulates secretion of fluid and electrolytes, enhances repair, and may minimize injury. To gain insight into the molecular processes influenced by KGF, we applied KGF to primary cultures of well-differentiated human airway epithelia and used microarray hybridization to assess the abundance of gene transcripts. Of 7,069 genes tested, KGF changed expression levels of 910. Earlier studies showed that KGF causes epithelial proliferation, and as expected, treatment altered expression of numerous genes involved in cell proliferation. We found that KGF stimulated transepithelial Cl−transport, but the number of cystic fibrosis (CF) transmembrane conductance regulator (CFTR) transcripts fell. Although transcripts for ClC-1 and ClC-7 Cl−channels increased, KGF failed to augment transepithelial Cl−transport in CF epithelia, suggesting that KGF-stimulated Cl−transport in differentiated airway epithelia depends on the CFTR Cl−channel. Interestingly, KGF decreased transcripts for many interferon (IFN)-induced genes. IFN causes trafficking of Stat dimers to the nucleus, where they activate transcription of IFN-induced genes. We found that KGF prevented the IFN-stimulated trafficking of Stat1 from the cytosol to the nucleus, suggesting a molecular mechanism for KGF-mediated suppression of the IFN-signaling pathway. These results suggest that in addition to stimulating proliferation and repair of damaged airway epithelia, KGF stimulates Cl−transport and may dampen the response of epithelial cells to inflammatory mediators.

scholarly journals In Vitro Modeling of Human Bocavirus 1 Infection of Polarized Primary Human Airway Epithelia

2013 ◽  
Vol 87 (7) ◽  
pp. 4097-4102 ◽  
Author(s):  
X. Deng ◽  
Z. Yan ◽  
Y. Luo ◽  
J. Xu ◽  
F. Cheng ◽  
...  
Keyword(s):  
Human Bocavirus ◽  
Airway Epithelia ◽  
In Vitro Modeling ◽  
Human Airway ◽  
Human Airway Epithelia
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