Gelatin-Based Microcarriers as Embryonic Stem Cell Delivery System in Bone Tissue Engineering:  An in-Vitro Study

Abstract

Mouse embryonic stem cells were cultured on commercially available biodegradable macroporous microcarriers. A culture period of 1-2 weeks was needed to colonize the microcarriers. Embryonic stem cells retained their pluripotency for up to 14 days when cultured in medium supplemented with leukemia inhibitory factor. Replacing this medium by differentiation medium for 2 weeks initiated osteogenic differentiation. Encapsulation of the cell-loaded microcarriers in photopolymerizable polymers (methacrylate-endcapped poly-D,L-lactide-co-caprolactone), triacetin/hydroxyethylmethacrylate (HEMA) as solvent and with/without gelatin as porogen, resulted in a homogeneous distribution of the microcarriers in the polymer. As observed by transmission electron microscopy, viability of the cells was optimal when gelatin was omitted and when using triacetin instead of HEMA.

Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells

“…Commercially available microcarriers-such as Cultisphere S, Cytodex 3, Solohill carriers and Hillex II have been widely used for the expansion of ESC, with Cytodex 3 being the most common. The speed used for mouse ESC, varies between 40 and 70 RPM (Fok and Zandstra 2005;Abranches et al 2007;Alfred et al 2011;Fernandes et al 2007;Marinho et al 2010;Storm et al 2010;Tielens et al 2007), while the range for human ESC lies within 24-80 RPM (Storm et al 2010;Chen et al 2011;Fernandes et al 2009;Kehoe et al 2010;Leung et al 2011;Lock and Tzanakakis 2009;Marinho et al 2013;Nie et al 2009;Oh et al 2009;Phillips et al 2008;Serra et al 2010). iPSC are by nature, delicate, making their large scale culture in spinner flasks using microcarriers a much more difficult proposition.…”

Skeletal tissue engineering using embryonic stem cells

“…ESCs could also be seeded and proliferated on gelatin-based microcarriers in a spinner flask. When proliferation medium was changed to osteogenic medium, indications of osteogenic differentiation were observed [158].…”

Evaluation of an Injectable, Photopolymerizable, and Three-Dimensional Scaffold Based on Methacrylate-Endcapped Poly(D,L-Lactide-co-ɛ-Caprolactone) Combined with Autologous Mesenchymal Stem Cells in a Goat Tibial Unicortical Defect Model

“…[7][8][9] Composition of the scaffolds was selected based on the results previously reported. 8,9 No extra porogen was included in the composition of the scaffolds.…”

“…[7][8][9] Composition of the scaffolds was selected based on the results previously reported. 8,9 No extra porogen was included in the composition of the scaffolds. Preceding experiments showed that porosity could be induced by the leaching of the plastizer and as a result of the degradation of the polyester and the CultiSpher microcarrier particles.…”

“…However, viability of the cells was estimated by transmission electronic microscopy to be optimal when gelatine was omitted, and triacetin was added instead of HEMA. 8 The objective of the present study was to evaluate the effect of mesenchymal stem cells loaded on microcarriers in combination with the crosslinkable methacrylate-endcapped porous polymer scaffold including a-TCP on bone regeneration in a unicortical tibial defect experimental design in a goat model. We firstly hypothesized that the implanted cells would easily survive and proliferate, and would demonstrate bone-forming characteristics inside the polymer scaffold.…”