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Proceedings Paper

Internal structures of scaffold-free 3D cell cultures visualized by synchrotron radiation-based micro-computed tomography
Author(s): Belma Saldamli; Julia Herzen; Felix Beckmann; Jutta Tübel; Johannes Schauwecker; Rainer Burgkart; Philipp Jürgens; Hans-Florian Zeilhofer; Robert Sader; Bert Müller
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Paper Abstract

Recently the importance of the third dimension in cell biology has been better understood, resulting in a re-orientation towards three-dimensional (3D) cultivation. Yet adequate tools for their morphological characterization have to be established. Synchrotron radiation-based micro computed tomography (SRμCT) allows visualizing such biological systems with almost isotropic micrometer resolution, non-destructively. We have applied SRμCT for studying the internal morphology of human osteoblast-derived, scaffold-free 3D cultures, termed histoids. Primary human osteoblasts, isolated from femoral neck spongy bone, were grown as 2D culture in non-mineralizing osteogenic medium until a rather thick, multi-cellular membrane was formed. This delicate system was intentionally released to randomly fold itself. The folded cell cultures were grown to histoids of cubic milli- or centimeter size in various combinations of mineralizing and non-mineralizing osteogenic medium for a total period of minimum 56 weeks. The SRμCT-measurements were performed in the absorption contrast mode at the beamlines BW 2 and W 2 (HASYLAB at DESY, Hamburg, Germany), operated by the GKSS-Research Center. To investigate the entire volume of interest several scans were performed under identical conditions and registered to obtain one single dataset of each sample. The histoids grown under different conditions exhibit similar external morphology of globular or ovoid shape. The SRμCT-examination revealed the distinctly different morphological structures inside the histoids. One obtains details of the histoids that permit to identify and select the most promising slices for subsequent histological characterization.

Paper Details

Date Published: 18 September 2008
PDF: 11 pages
Proc. SPIE 7078, Developments in X-Ray Tomography VI, 70781X (18 September 2008); doi: 10.1117/12.795162
Show Author Affiliations
Belma Saldamli, Johann Wolfgang Goethe-Univ. Frankfurt am Main (Germany)
Univ. Hospital Basel (Switzerland)
Julia Herzen, GKSS-Research Ctr. Geesthacht (Germany)
Felix Beckmann, GKSS-Research Ctr. Geesthacht (Germany)
Jutta Tübel, Technische Univ. München (Germany)
Johannes Schauwecker, Technische Univ. München (Germany)
Rainer Burgkart, Technische Univ. München (Germany)
Philipp Jürgens, Univ. Basel (Switzerland)
Hans-Florian Zeilhofer, Univ. Basel (Switzerland)
Robert Sader, Johann Wolfgang Goethe-Univ. Frankfurt am Main (Germany)
Bert Müller, Univ. Basel (Switzerland)


Published in SPIE Proceedings Vol. 7078:
Developments in X-Ray Tomography VI
Stuart R. Stock, Editor(s)

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