Abstract
Easily assembled organotypic co-cultures have long been sought in medical research. In vitro tissue constructs with faithful representation of in vivo tissue characteristics are highly desirable for screening and characteristic assessment of a variety of tissue types. Cardiac tissue analogs are particularly sought after due to the phenotypic degradation and difficulty of culture of primary cardiac myocytes. This study utilized magnetic nanoparticles and primary cardiac myocytes in order to levitate and culture multicellular cardiac aggregates (MCAs). Cells were isolated from 2 day old Sprague Dawley rat hearts and subsequently two groups were incubated with either C1: 33 µL nanoshell/million cells or C2: 50 µL nanoshell/million cells. Varying numbers of cells for each concentration were cultured in a magnetic field in a 24 well plate and observed over a period of 12 days. Constructs generally formed spherical structures. Masson’s trichrome staining of a construct shows the presence of extracellular matrix protein, indicating the presence of functional fibroblasts. Many constructs exhibited noticeable contraction after 4 days of culture and continued contracting noticeably past day 9 of culture. Noticeable contractility indicates the presence of functional primary cardiac myocytes in culture. Phenotypic conservation of cardiac cells was ascertained using IHC staining by α-actinin and collagen. CD31 and fibrinogen were probed in order to assess localization of fibroblasts and endothelial cells. The study verifies a protocol for the use of magnetic levitation in order to rapidly assemble 3D cardiac like tissue with phenotypic and functional stability.
Highlights
Three-dimensional (3D) cell culture systems have been a critical area of biomedical research for decades
The technique presented in this paper was used to successfully generate hundreds of multicellular cardiac aggregates (MCAs) with relative ease
The MCAs presented with similar geometry and formed with a 100% success rate
Summary
Three-dimensional (3D) cell culture systems have been a critical area of biomedical research for decades. Organotypic culture systems, have been a central area of concern in biomedical research for decades. Streamlining materials and simplifying the model can reduce such costs and make an organotypic culture system more palatable for product development. A 3D culture model must provide useful information. Towards this end, a highly representative environment is desirable. Scaffolds tend to limit the amount of cell-cell connectivity possible. This is important when considering models involving cells like cardiac myocytes that contain gap junctions that are essential to the functionality of said cells
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