Acute cytotoxicity and apoptotic effects after l-Pam exposure in different cocultures of the proximal and distal respiratory system

Pohl, Christine; Hofmann, Hélène; Moisch, Michaela; Papritz, Mirko; Hermanns, Maria Iris; Dei-Anang, Jasmin; Mayer, Eckhard; Kehe, Kai; Kirkpatrick, Charles James

doi:10.1016/j.jbiotec.2009.12.007

Cited by 11 publications

(9 citation statements)

References 24 publications

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“…The simplest co-culture models use differentiated or undifferentiated airway epithelial cells in the apical compartment of semi-permeable filter support units and other cell types in the basal compartment. Adherent cells such as fibroblasts or endothelial cells can also be cultured in close contact with the epithelial cells on the under-surface of the filter insert (Pohl et al, 2010). A commercially available co-culture model that allows direct contact between the differentiated airway epithelium and a fibroblastcontaining subepithelial matrix is also available (EpiAirwayFT, MatTek).…”

Section: Co-culture and Pathophysiological Modelsmentioning

confidence: 99%

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

Gordon

Daneshian

Bouwstra

et al. 2015

ALTEX

124

103

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SummaryModels of the outer epithelia of the human body -namely the skin, the intestine and the lung -have found valid applications in both research and industrial settings as attractive alternatives to animal testing. A variety of approaches to model these barriers are currently employed in such fields, ranging from the utilization of ex vivo tissue to reconstructed in vitro models, and further to chip-based technologies, synthetic membrane systems and, of increasing current interest, in silico modeling approaches. An international group of experts in the field of epithelial barriers was convened from academia, industry and regulatory bodies to present both the current state of the art of non-animal models of the skin, intestinal and pulmonary barriers in their various fields of application, and to discuss research-based, industry-driven and regulatory-relevant future directions for both the development of new models and the refinement of existing test methods. Issues of model relevance and preference, validation and standardization, acceptance, and the need for simplicity versus complexity were focal themes of the discussions. The outcomes of workshop presentations and discussions, in relation to both current status and future directions in the utilization and development of epithelial barrier models, are presented by the attending experts in the current report.

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Section: Co-culture and Pathophysiological Modelsmentioning

confidence: 99%

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

Gordon

Daneshian

Bouwstra

et al. 2015

ALTEX

124

103

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“…cup model for vapor exposure in place of topical mustard exposures. 20 In previous studies using topical exposure to the NM derivative 4-[bis(2chloroethyl)amino]-l-phenylalanine (melphalan; l-Pam), Pohl et al 27 demonstrated decreased viability and decreased TEER measurements in a differentiated airway epithelial culture. Consistent with prior topical exposures, our results demonstrate increased LDH expression and epithelial cellular detachment in airway epithelial cultures at 24 h postexposure after a 1-h 50 μM NM exposure.…”

Section: Discussionmentioning

confidence: 99%

“…To the best of our knowledge, this is also the first study to evaluate the effects of NM exposure on the coculture of primary AECs with primary PAECs. Using transformed cell lines, Pohl et al 27 evaluated the effects of topical melphalan on a coculture alveolar model of a lung adenocarcinoma cell line NCI H441 with an angiosarcoma cell line Iso-Has-1. This culture model is most reflective of the distal lung parenchyma and less reflective of airway pathology.…”

Section: Discussionmentioning

confidence: 99%

“…This design provided greater barrier protection to the underlying endothelial layer than a single monolayer culture used in the prior study. In Pohl et al, 27 endothelial cells were cultured directly on the basolateral surface of the permeable cell insert. Thus, the current study is likely more reflective of the airway compartment, considering that PAECs do not grow directly on the basement membrane of the airway, while the prior study may be a more reflective model of the distal lung parenchyma, where epithelial and endothelial cells grow adjacent to one another at the alveolar-capillary interface.…”

Section: Discussionmentioning

confidence: 99%

“…We adapted a vapor cup model for vapor exposure in place of topical mustard exposures 20 . In previous studies using topical exposure to the NM derivative 4‐[bis(2‐chloroethyl)amino]‐ l ‐phenylalanine (melphalan; l ‐Pam), Pohl et al 27 . demonstrated decreased viability and decreased TEER measurements in a differentiated airway epithelial culture.…”

Section: Discussionmentioning

confidence: 99%

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Acute cytotoxicity and increased vascular endothelial growth factor after in vitro nitrogen mustard vapor exposure

McGraw

Kim

White

et al. 2020

Annals of the New York Academy of Sciences

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Nitrogen mustard (NM) is a highly toxic alkylating agent. Inhalation exposure can cause acute and chronic lung injury. This study's aims were to develop an in vitro coculture model of mustard-induced airway injury and to identify growth factors contributing to airway pathology. Primary human bronchial epithelial cells cultured with pulmonary endothelial cells were exposed to NM (25, 50, 100, 250, or 500 µM) or PBS (control) for 1 hour. Lactate dehydrogenase (LDH) and transepithelial electrical resistance (TEER) were measured before and 24 h after NM exposure. Fixed cultures were stained for hematoxylin and eosin or live/dead staining. Culture media were analyzed for 11 growth factors. A 1-h vapor exposure to greater than or equal to 50 µM NM increased supernatant LDH, decreased TEER, and caused airway epithelial cell detachment. Endothelial cell death occurred at 500 µM NM. Vascular endothelial growth factor A (VEGF-A) and placental growth factor (PlGF) expression increased in 500 µM NM−exposed cultures compared with PBS-exposed control cultures. NM vapor exposure causes differential cytotoxicity to airway epithelial and endothelial injury in culture. Increased VEGF-A and PlGF expression occurred acutely in airway cocultures. Future studies are required to validate the role of VEGF signaling in mustardinduced airway pathology.

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3.15 Cell Culture Systems for Studying Biomaterial Interactions With Biological Barriers ☆

Hermanns¹,

Freese²,

Anspach³

et al. 2017

Comprehensive Biomaterials II

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Acute cytotoxicity and apoptotic effects after l-Pam exposure in different cocultures of the proximal and distal respiratory system

Cited by 11 publications

References 24 publications

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

Acute cytotoxicity and increased vascular endothelial growth factor after in vitro nitrogen mustard vapor exposure

3.15 Cell Culture Systems for Studying Biomaterial Interactions With Biological Barriers ☆

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