Three-Dimensional In Vitro Reaggregates of Embryonic Cardiomyocytes: A Potential Model System for Monitoring Effects of Bioactive Agents

Bartholomä, P.; Gorjup, Erwin; Monz, D.; Reininger-Mack, A.; Thielecke, Hagen; Robitzki, Andrea A.

doi:10.1177/1087057105280070

Cited by 23 publications

(15 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…13 A suitable 3D substrate to support organizational cultures has been applied to liver cells, cardiac cells, and solid tumors to attain morphological structure and maintain the integrity of intercellular signaling mechanisms. 15,[28][29][30] Collagen, a major constituent of the ECM, 31 has consistently been shown to support the development of the embryoid bodies. The microfilaments supporting structural collagen likely enable the cells to grow and organize naturally into an in vivo-like morphology.…”

Section: Discussionmentioning

confidence: 99%

Induction of Ciliated Cells from Avian Embryonic Stem Cells Using Three-Dimensional Matrix

Wang

Wong

Mao

2010

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

We have devised a simple three-dimensional (3D) tissue-culturing method to induce ciliogenesis from avian embryonic stem (ES) cells by using avian fertilized eggs. Unlike the previous reported techniques, this method does not require trypsinization, which would reduce the viability of the cells; it also does not require an airliquid interface to induce ciliogenesis and to maintain the growth of the induced ciliated cells. ES cells seeded and attached on this collagen-coated chitosan 3D gel grew spontaneously and robustly. Following 2 weeks in culture with inhibition of embryoid body formation, cells with noticeable and vigorous beating cilia were observed. We measured the ciliary beat frequencies of these ES-differentiated ciliated cells for 40 days. These results were consistent with all reported measurements made for other species of ciliated cells, including human, from our previous study. These data imply that the cilia of these ES-derived ciliated cells, beating at their intrinsic basal autorhythmic rate, preserve the integrity of the regulatory mechanisms of ciliary beat frequency. In conclusion, we have shown that ES cells cultured in a 3D tissue-engineered scaffold is a promising approach for developing an in vitro cell model that closely mimics the in vivo ciliated cell natural milieu. This cell model can potentially be the source of ciliated cells for cell-based high-throughput screening and discovery of pulmonary drugs.

show abstract

Section: Discussionmentioning

confidence: 99%

Induction of Ciliated Cells from Avian Embryonic Stem Cells Using Three-Dimensional Matrix

Wang

Wong

Mao

2010

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

show abstract

“…9,10 These processes have led to the formation of several different micro-tissue spheroids including bile canniculi 11 and beating cardiomyocytes. 12,13 LaMarca et al created 3D trophoblast aggregates of the SGHPL-4 cell line and demonstrated that the 3D configuration induced the invasive properties of these cells. This important study illustrated that trophoblast cells cultured in 3D have functional behavior strikingly different from those in monolayer.…”

Section: Discussionmentioning

confidence: 99%

Bioengineering Anembryonic Human Trophoblast Vesicles

et al. 2011

View full text Add to dashboard Cite

Introduction: Trophoblast cells in vivo form a 3-dimensional structure that promotes complex cell-to-cell interactions that cannot be studied with traditional monolayer culture. We describe a 3-dimensional trophoblast bioreactor to study cellular interactions. Methods: Nonadhesive agarose hydrogels were cast from molds using computer-assisted prototyping. Trophoblast cells were seeded into the gels for 10 days. Morphology, viability, and vesicle behavior were assessed. Results: Trophoblast cells formed uniform spheroids. Serial sectioning on days 3, 7, and 10 revealed central vacuolization with a consistent outer rim 12.3-m thick. The vesicle configuration has been confirmed with confocal imaging. Electron Microscopic (EM) imaging revealed its ultrastructure. The vesicles migrate across a fibronectin-coated surface and invaded basement membrane. Conclusions: Trophoblast cells cultured in a novel substrate-free 3-dimensional system form trophoblast vesicles. This new cell culture technique allows us to better study placental cell-to-cell interactions with the potential of forming microtissues. Keywords trophoblast cell, blastocyst, 3-dimensional cultureHuman cellular differentiation begins 5 days postconception in the blastocyst that is composed of inner cell mass (ICM) cells and trophoblast cells. The ICM ultimately forms the embryo. 1The trophoblast cells, the focus of this research, further differentiate into the cells required to attach the embryo to the uterine wall, invade the endometrium and the maternal vasculature, 2 and form the maternal-fetal interface and the hormone producing placental cells.3 Surprisingly, little is known about the molecular mechanisms that guide the process of early differentiation.A wealth of information about placental cell differentiation has been gained from traditional 2-dimensional (2D) monolayer culture. However, traditional 2D cell culture does not allow the study of cell-to-cell interactions. Furthermore, monolayer culture prevents analysis of the gene programs required to form complex cellular structures such as trophoblast vesicles and the placenta. We have recently developed a method to culture trophoblast cells into self-assembled symmetric 3D vesicles using nonadhesive micromold bioreactors. 4 MethodsWax molds were designed in computer-assisted design (Solid Works Corporation, Concord, Massachusetts) and produced with a ThermoJet rapid prototyping machine (3D Systems Corporation, Valencia, California) as described by Dean et al. 3 Each mold contained a matrix of 822 cylindrical bottom pegs 200 m in diameter. Agarose gel bioreactors were cast directly from wax molds. Aliquots of 2 g Ultra Pure Agarose (American Bioanalytical, Natick, Massachusetts) were autoclaved as a powder, 200 mL of sterile dH 2 O was added and the agarose was dissolved by heating and mixing on a hot plate. The solution was cooled and approximately 2.75 mL was pipetted into each wax mold in a sterile dish. After setting, the bioreactors were separated from the mold using a spatula, tr...

show abstract

“…Establishment of spheroids followed a modified protocol of Bartholomä et al [2005]. Fertile chicken eggs (white Leghorn) were incubated at 37 8C.…”

Section: Cell Culturementioning

confidence: 99%

Electromagnetic exposure of scaffold-free three-dimensional cell culture systems

Daus

Goldhammer

Layer

et al. 2011

Bioelectromagnetics

View full text Add to dashboard Cite

In recent years, a number of in vitro studies have reported on the possible athermal effects of electromagnetic exposure on biological tissue. Typically, this kind of study is performed on monolayers of primary cells or cell lines. However, two-dimensional cell layer systems lack physiological relevance since cells in vivo are organized in a three-dimensional (3D) architecture. In monolayer studies, cell-cell and cell-ECM interactions obviously differ from live tissue and scale-ups of experimental results to in vivo systems should be considered carefully. To overcome this problem, we used a scaffold-free 3D cell culture system, suitable for the exploration of electrophysiological effects due to electromagnetic fields (EMF) at 900 MHz. Dissociated cardiac myocytes were reaggregated into cellular spheres by constant rotation, and non-invasive extracellular recordings of these so-called spheroids were performed with microelectrode arrays (MEA). In this study, 3D cell culture systems were exposed to pulsed EMFs in a stripline setup. We found that inhomogeneities in the EMF due to electrodes and conducting lines of the MEA chip had only a minor influence on the field distribution in the spheroid if the exposure parameters were chosen carefully.

show abstract

Three-Dimensional In Vitro Reaggregates of Embryonic Cardiomyocytes: A Potential Model System for Monitoring Effects of Bioactive Agents

Cited by 23 publications

References 38 publications

Induction of Ciliated Cells from Avian Embryonic Stem Cells Using Three-Dimensional Matrix

Induction of Ciliated Cells from Avian Embryonic Stem Cells Using Three-Dimensional Matrix

Bioengineering Anembryonic Human Trophoblast Vesicles

Electromagnetic exposure of scaffold-free three-dimensional cell culture systems

Contact Info

Product

Resources

About