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

Processes

Ioannis Papantoniou¹  Maarten Sonnaert¹  Toon Lambrechts¹  Jean-Marie Aerts¹  Lies Geris¹  Frank P. Luyten¹ 

[1] Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, Onderwijs en Navorsing 1 (+8), Herestraat 49-PB813, B-3000 Leuven, Belgium; E-Mails:

关键词: perfusion bioreactor;    osteogenic differentiation;    gene expression;    DoE analysis;    tissue engineering;    biomanufacturing;   

DOI  :  10.3390/pr2030639

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.