Three-dimensional growth patterns of various human tumor cell lines in simulated microgravity of a NASA bioreactor

Abstract

Growth patterns of a number of human tumor cell lines that from three-dimensional structures of various architectures when cultured without carrier beads in a NASA rotary cell culture system are described and illustrated. The culture system, which was designed to mimic microgravity, maintained cells in suspension under very low-shear stress throughout culture. Spheroid (particulate) production occurred within a few hours after culture was started, and spheroids increased in size by cell division and fusion of small spheroids, usually stabilizing at a spheroid diameter of about 0.5 mm. Architecture of spheroids varied with cell type. Cellular interactions that occurred in spheroids resulted in conformation and shape changes of cells, and some cell lines produced complex, epithelial-like architectures. Expression of the cell adhesion molecules, CD44 and E cadherin, was upregulated in the three-dimensional constructs. Coculture of fibroblast spheroids with PC3 prostate cancer cells induced tenascin expression by the fibroblasts underlying the adherent prostate epithelial cells. Invasion of the fibroblast spheroids by the malignant epithelium was also demonstrated.

Advances in multicellular spheroids formation

“…Many approaches have been developed to fabricate MCSs, such as dispersed cells (referred to as 'matrix-free' herein), cells embedded in hydrogel matrix (referred to as 'matrix-based' herein) and microfluidic platforms as shown in figure 4. The matrix-free approach has been commonly used to produce MCSs, such as suspension culture in non-adhesive surfaces [18,50 -52], hanging drops [53,54], spinner flasks or rotational bioreactors [55][56][57] and external-force-driven MCS aggregation [21,58 -62]. These methods have been detailed and reviewed in previous papers [24,63,64].…”

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

“…Ingram and co-workers manufactured a new rotary cell culture system, termed as NASA, that allows a dynamic control over the spheroids culture conditions. [48] The NASA bioreactor allows the precise control over the pH of the culture medium and the concentration of nutrients and gases (e.g., glucose and oxygen, respectively). [49,50] However, the use of these complex systems increases the expenses associated to spheroids production and some of the shear forces drawbacks are not completely surpassed.…”

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

“…Several malignant glioma cell lines were introduced in the NASA rotary cell culture system, forming spheroids with an increasing expression of the adhesion molecule CD44 over the time in culture [260]. An exhaustive study on brain tumor aggregates generated using the 3D RCCS TM has been presented recently by Smith and collaborators [261]. Panchalingam and coworkers [262] developed bioreactor protocols for expanding cancer stem cells from human glioblastoma specimens.…”