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Processes 2014, 2(3), 639-657; doi:10.3390/pr2030639

Analysis of Gene Expression Signatures for Osteogenic 3D Perfusion-Bioreactor Cell Cultures Based on a Multifactorial DoE Approach

1
Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, Onderwijs en Navorsing 1 (+8), Herestraat 49-PB813, B-3000 Leuven, Belgium
2
Skeletal Biology and Engineering Research Center, KU Leuven, Onderwijs en Navorsing 1 (+8), Herestraat 49-PB813, B-3000 Leuven, Belgium
3
Department of Materials Engineering, KU Leuven, Kasteelpark Arenberg 44-PB 2450, B-3001 Heverlee, Belgium
4
Division Measure, Model & Manage Bioresponses (M3-BIORES), KU Leuven, Kasteelpark Arenberg 30, B-3001 Leuven, Belgium
5
Biomechanics Research Unit, Universite de Liege, Chemin des Chevreuils 1-BAT 52/3, B-4000 Liege, Belgium
*
Author to whom correspondence should be addressed.
Received: 14 February 2014 / Revised: 9 May 2014 / Accepted: 9 July 2014 / Published: 8 August 2014
(This article belongs to the Special Issue Design of Bioreactor Systems for Tissue Engineering)
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Abstract

The use of multifactorial design of experiments (DoE) in tissue engineering bioprocess development will contribute to the robust manufacturing of tissue engineered constructs by linking their quality characteristics to bioprocess operating parameters. In this work, perfusion bioreactors were used for the in vitro culture and osteogenic differentiation of human periosteum-derived cells (hPDCs) seeded on three-dimensional titanium (Ti) alloy scaffolds. A CaP-supplemented medium was used to induce differentiation of the cultured hPDCs. A two-level, three-factor fractional factorial design was employed to evaluate a range of bioreactor operating conditions by changing the levels of the following parameters: flow rate (0.5–2 mL/min), cell culture duration (7–21 days) and cell seeding density (1.5 × 103–3 × 103 cells/cm2). This approach allowed for evaluating the individual impact of the aforementioned process parameters upon a range of genes that are related to the osteogenic lineage, such as collagen type I, alkaline phosphatase, osterix, osteopontin and osteocalcin. Furthermore, by overlaying gene-specific response surfaces, an integrated operating process space was highlighted within which predetermined values of the six genes of interest (i.e., gene signature) could be minimally met over the course of the bioreactor culture time. View Full-Text
Keywords: perfusion bioreactor; osteogenic differentiation; gene expression; DoE analysis; tissue engineering; biomanufacturing perfusion bioreactor; osteogenic differentiation; gene expression; DoE analysis; tissue engineering; biomanufacturing
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Papantoniou, I.; Sonnaert, M.; Lambrechts, T.; Aerts, J.-M.; Geris, L.; Luyten, F.P.; Schrooten, J. Analysis of Gene Expression Signatures for Osteogenic 3D Perfusion-Bioreactor Cell Cultures Based on a Multifactorial DoE Approach. Processes 2014, 2, 639-657.

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