Human tissue-engineered bone produced in clinically relevant amounts using a semi-automated perfusion bioreactor system: a preliminary study

被引:34
作者
Janssen, F. W. [1 ,2 ]
van Dijkhuizen-Radersma, R. [2 ]
Van Oorschot, A. [3 ]
Oostra, J. [4 ]
de Bruijn, J. D. [2 ,5 ]
Van Blitterswijk, C. A. [1 ]
机构
[1] Univ Twente, Inst Biomed Technol, NL-3720 AB Bilthoven, Netherlands
[2] Xpand Biotechnol BV, NL-3723 MB Bilthoven, Netherlands
[3] IsoTis SA, NL-3723 MB Bilthoven, Netherlands
[4] Applikon Dependable Instruments BV, NL-3100 AC Schiedam, Netherlands
[5] Queen Mary Univ London, London, England
关键词
bioreactor; tissue engineering; bone; human bone marrow stem cells; perfusion; online measurement; in vivo; MARROW STROMAL CELLS; MESENCHYMAL STEM-CELLS; MINERALIZED MATRIX DEPOSITION; IN-VIVO; SPINE FUSION; CULTURE; VITRO; OSTEOGENESIS; MULTIPOTENCY; OSTEOBLASTS;
D O I
10.1002/term.197
中图分类号
Q813 [细胞工程];
学科分类号
摘要
The aim of this study was to evaluate a semi-automated perfusion bioreactor system for the production of clinically relevant amounts of human tissue-engineered bone. Human bone marrow stromal cells (hBMSCs) of eight donors were dynamically seeded and proliferated in a perfusion bioreactor system in clinically relevant volumes (10 cm(3)) of macroporous biphasic calcium phosphate scaffolds (BCP particles, 2-6 mm). Cell load and distribution were shown using methylene blue staining. MTT staining was used to demonstrate viability of the present cells. After 20 days of cultivation, the particles were covered with a homogeneous layer of viable cells. Online oxygen measurements confirmed the proliferation of hBMSCs in the bioreactor. After 20 days of cultivation, the hybrid constructs became interconnected and a dense layer of extracellular matrix was present, as visualized by scanning electron microscopy (SEM). Furthermore, the hBMSCs showed differentiation towards the osteogenic lineage as was indicated by collagen type I production and alkaline phosphatase (ALP) expression. We observed no significant differences in osteogenic gene expression profiles between static and dynamic conditions like ALP, BMP2, Id1, Id2, Smad6, collagen type 1, osteocalcin, osteonectin and S100A4. For the donors that showed bone formation, dynamically cultured hybrid constructs showed the same amount of bone as the statically cultured hybrid constructs. Based on these results, we conclude that a semi-automated perfusion bioreactor system is capable of producing clinically relevant and viable amounts of human tissue-engineered bone that exhibit bone-forming potential after implantation in nude mice. Copyright (C) 2009 John Wiley & Sons, Ltd.
引用
收藏
页码:12 / 24
页数:13
相关论文
共 53 条
[1]   Cell culture: Biology's new dimension [J].
Abbott, A .
NATURE, 2003, 424 (6951) :870-872
[2]   LacZ and interleukin-3 expression in vivo after retroviral transduction of marrow-derived human osteogenic mesenchymal progenitors [J].
Allay, JA ;
Dennis, JE ;
Haynesworth, SE ;
Majumdar, MK ;
Clapp, DW ;
Shultz, LD ;
Caplan, AI ;
Gerson, SL .
HUMAN GENE THERAPY, 1997, 8 (12) :1417-1427
[3]  
Aubin JE, 1998, J CELL BIOCHEM, P73, DOI 10.1002/(SICI)1097-4644(1998)72:30/31+<73::AID-JCB11>3.0.CO
[4]  
2-L
[5]  
BAB I, 1988, BONE MINER, V4, P373
[6]   Design of a flow perfusion bioreactor system for bone tissue-engineering applications [J].
Bancroft, GN ;
Sikavitsas, VI ;
Mikos, AG .
TISSUE ENGINEERING, 2003, 9 (03) :549-554
[7]   Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteloblasts in a dose-dependent manner [J].
Bancroft, GN ;
Sikavitsast, VI ;
van den Dolder, J ;
Sheffield, TL ;
Ambrose, CG ;
Jansen, JA ;
Mikos, AG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (20) :12600-12605
[8]   Proliferation kinetics and differentiation potential of ex vivo expanded human bone marrow stromal cells: Implications for their use in cell therapy [J].
Banfi, A ;
Muraglia, A ;
Dozin, B ;
Mastrogiacomo, M ;
Cancedda, R ;
Quarto, R .
EXPERIMENTAL HEMATOLOGY, 2000, 28 (06) :707-715
[9]  
Bartrons R, 2007, J BIOENERG BIOMEMBR, V39, P223, DOI [10.1007/s10863-007-9080-3, 10.1007/S10863-007-9080-3]
[10]   Marrow stromal stem cells [J].
Bianco, P ;
Robey, PG .
JOURNAL OF CLINICAL INVESTIGATION, 2000, 105 (12) :1663-1668