International Review of Cell and Molecular Biology (eBook)

Kwang W. Jeon (Herausgeber)

eBook Download: PDF | EPUB

2015 | 1. Auflage
300 Seiten
Elsevier Science (Verlag)
978-0-12-802476-8 (ISBN)

Infos

International Review of Cell and Molecular Biology presents comprehensive reviews and current advances in cell and molecular biology. Articles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. The series has a worldwide readership, maintaining a high standard by publishing invited articles on important and timely topics authored by prominent cell and molecular biologists. - Includes insights from the foremost scientists in the field, with specific discussions of the current state of research on gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and more - Provides comprehensive reviews and current advances - Presents a wide range of perspectives on specific subjects - Valuable reference material for advanced undergraduates, graduate students, and professional scientists

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2. 2D Cell Culture

In many experimental setups, the creation of a functional host cell surface is sufficient to study initial interactions with bacterial pathogens such as adherence, invasion, and induction of signal transduction processes. For decades of years 2D cell monolayers grown on solid, impermeable plastic or glass surfaces have been applied as simple and cost-effective strategy to analyze principle mechanisms of bacterial–host cell interactions. Numerous in vitro cell culture systems have been confirmed as suitable to provide information about specific bacterial virulence factors involved and also elucidated the induction of many fold intracellular processes, such as signaling cascades and cytoskeletal rearrangements on the host side (Table 1).

Depending on the physiological niche of the bacteria, pulmonary cells are cultivated and infected with typical lung pathogens like Legionella pneumophila and Streptococcus pneumoniae; gastrointestinal cells are used for infection studies with Helicobacter and Salmonella, and skin fibroblasts are chosen for infection with causative agents of wound infections like staphylococci—just giving a few examples. These studies generated impressive transmission electron microscopic pictures and opened up the field for more detailed cell culture infection analyses. Several immunofluorescence staining procedures have been developed, which can be applied after infection of eukaryotic cell monolayers with pathogenic bacteria. These procedures enable a microscopic visualization and also a differential quantification of adhered and internalized bacteria (Bergmann et al., 2009, 2013; Elm et al., 2004; Jensch et al., 2010; Lüttge et al., 2012; Agarwal et al., 2013, 2014; Pracht et al., 2005; Amelung et al., 2011; Nerlich et al., 2009; Rohde and Chhatwal, 2013). In addition, fluorescence staining of the actin cytoskeleton also visualized radical morphological changes of the eukaryotic cells, e.g., induced by streptococcal adherence (Bergmann et al., 2009). Complex responses of host immune systems are also studied using suspension cultures with prepared and differentiated macrophages or other cells derived from the lymphoid tissues.

Table 1

Representative examples of bacterial pathogens analyzed in different cell culture models as highlighted in this review

Single cell type monolayer

Macrophages

Helicobacter pylori

Wang et al., 2009

Macrophages, Dictyostelium discoideum

Legionella pneumophila

Steinert et al. (1994), Allombert et al. (2014), Steinert et al. (2000), Shevchuk and Steinert (2009), and Skriwan et al. (2002)

D. discoideum

Mycobacterium marinum

Meng et al. (2014)

Dendritic cells

Streptococcus pneumoniae

Rosendahl et al. (2013)

Epithelial and endothelial cells

S. pneumoniae

Steinford et al. (1989), Jensch et al. (2010), Bergmann et al. (2009, 2013), Lüttge et al. (2012), Agarwal et al. (2013, 2014), Pracht et al. (2005), and Elm et al. (2004)

Endothelial cells

Streptococcus pyogenes

Amelung et al. (2011) and Nerlich et al. (2009)

2D coculture model

Bilayer model (epithelium and endothelium)

Neisseria meningitides

Birkness et al. (1995)

Blood–brain barrier model

Escherichia coli

Huang et al. (1995, 2000) and Kim (2000)

Streptococcus agalactiae

Nizet et al. (1997)

Listeria monocytogenes

Greiffenberg et al. (1998)

Citrobacter feundii

Badger et al. (1999)

S. pneumoniae

Ring et al. (1998) and Untucht et al. (2011)

Blood–cerebrospinal fluid model

N. meningitides

Steinmann et al. (2013)

Lung coculture model

Pseudomonas aeruginosa, Klebsiella pneumoniae, E. coli

Hurley et al. (2004)

3D culture model with scaffolds

Transwell system with ECM

Chlamydia trachomatis

Igietseme et al. (1994), Kane and Byrne (1998), Kane et al. (1999), and Dessus-Babus et al. (2002)

L. pneumophila

Skriwan et al. (2002)

L. monocytogenes

Cossart and Lecuit (1998)

Neisseria gonoorhoeae

Hopper et al. (2000)

S. pyogenes

Ochel et al. (2014)

Table Continued

Epithelial airway tissue model

S. pneumoniae

Fliegauf et al. (2013)

P. aeruginosa

Woodworth et al. (2008)

Rotary wall vessel culture

A549 lung epithelial cells

Francisella tularensis

David et al. (2014)

P. aeruginosa

Carterson et al. (2005)

Cells on collagen-coated microcarrier beads

C. trachomatis

Guseva et al. (2007)

Human intestinal Int-407 cells

Salmonella enterica

Nickerson et al. (2001)

Organoids

Human ileocecal colorectal adenocarcinoma HCT-8

E. coli (EHEC/EPEC)

Carvalho et al. (2005)

Human skin equivalent

Acinetobacter baumannii

de Breij et al. (2010) and Breij et al. (2014)

Tissue explants

Lung tissue explants

Chlamydia pneumophila

Rupp et al. (2004)

L. pneumophila

Jäger et al. (2014) and Shevchuk et al. (2014)

S. pneumoniae

Szymanski et al. (2012)

Tonsil explants

S. pyogenes

Bell et al. (2012) and Abbot et al. (2007)

Microfluidic perfusion

HUVEC

Staphylococcus aureus

Pappelbaum et al. (2013)

This chapter will provide an overview about the broad spectrum of 2D cell culture models in infection biology. Beginning with the discussion of key aspects in using immortalized versus primary nonimmortalized eukaryotic monolayers in infection models, the second part is focused on protozoa-based models in infection biology. Climbing to the next level of cell culture complexity, the advantages of coculture models generated by simultaneous cultivation of two or more different cell types will be described. The cocultivation technique is used to regenerate tissue barriers and will be demonstrated by the examples of a transwell-based reconstruction of a blood–brain barrier (BBB) and a blood–cerebrospinal fluid barrier (BCSFB).

2.1. Culture of Immortalized Cell Lines versus Primary Cell Culture

Many valuable bacterial pathogenicity mechanisms have been elucidated using 2D monolayer cell cultures indicating that certain scientific questions can be answered and sometimes even require this kind of simplified cell culture technique. A risk of using 2D monolayer cell culture models is the loss or diminished expression of certain phenotypic characteristics that may mediate bacterial–host cell interactions. This loss of phenotype results in progressive alterations in biochemistry, function, and morphology and increases with every passage of the culture as the cells diverge from the source tissue phenotype (Shaw, 1996). Of special importance are immortalized human cell lines, which have been used extensively for the study of host–pathogen interactions. These stable cell lines combine the advantage of an indefinite life span allowing passaging of several hundred times with low or moderate culture requirements. However, these lines exhibit aberrant properties attributable to immortalization and artificial 2D growth conditions (Freshney, 2005).

Thus, several studies targeting the investigation of pathogen–cell interactions have been conducted with primary cells derived from different animals such as primary porcine endothelial and epithelial cells (Vanier et al., 2007;...

Produkt-Details

• Erscheint lt. Verlag: 16.11.2015
• Sprache: englisch
• Themenwelt: Naturwissenschaften ► Biologie ► Genetik / Molekularbiologie
• Themenwelt: Naturwissenschaften ► Biologie ► Zellbiologie
• Themenwelt: Technik
• ISBN-10: 0-12-802476-3 / 0128024763
• ISBN-13: 978-0-12-802476-8 / 9780128024768