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Encapsulation of adult human mesenchymal stem cells within collagen‐agarose microenvironments
Author(s) -
Batorsky Anna,
Liao Jiehong,
Lund Amanda W.,
Plopper George E.,
Stegemann Jan P.
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20614
Subject(s) - agarose , extracellular matrix , mesenchymal stem cell , cell encapsulation , fluorescence microscope , matrix (chemical analysis) , bead , tissue engineering , centrifugation , chemistry , materials science , biophysics , cell , microbiology and biotechnology , biomedical engineering , chromatography , fluorescence , biochemistry , biology , composite material , medicine , physics , quantum mechanics
Abstract Reliable control over the process of cell differentiation is a major challenge in moving stem cell‐based therapies forward. The composition of the extracellular matrix (ECM) is known to play an important role in modulating differentiation. We have developed a system to encapsulate adult human mesenchymal stem cells (hMSC) within spherical three‐dimensional (3D) microenvironments consisting of a defined mixture of collagen Type I and agarose polymers. These protein‐based beads were produced by emulsification of liquid hMSC‐matrix suspensions in a silicone fluid phase and subsequent gelation to form hydrogel beads, which were collected by centrifugation and placed in culture. Bead size and size distribution could be varied by changing the encapsulation parameters (impeller speed and blade separation), and beads in the range of 30–150 microns in diameter were reliably produced. Collagen concentrations up to 40% (wt/wt) could be incorporated into the bead matrix. Visible light and fluorescence microscopy confirmed that the collagen matrix was uniformly distributed throughout the beads. Cell viability post‐encapsulation was in the range of 75–90% for all bead formulations (similar to control slab gels) and remained at this level for 8 days in culture. Fluorescent staining of the actin cytoskeleton revealed that hMSC spreading increased with increasing collagen concentration. This system of producing 3D microenvironments of defined matrix composition therefore offers a way to control cell‐matrix interactions and thereby guide hMSC differentiation. The bead format allows the use of small amounts of matrix proteins, and such beads can potentially be used as a cell delivery vehicle in tissue repair applications. © 2005 Wiley Periodicals, Inc.