Bioreactor design for successive culture of anchorage-dependent cells operated in an automated manner.

A novel bioreactor system was designed to perform a series of batchwise cultures of anchorage-dependent cells by means of automated operations of medium change and passage for cell transfer. The experimental data on contamination frequency ensured the biological cleanliness in the bioreactor system, which facilitated the operations in a closed environment, as compared with that in flask culture system with manual handlings. In addition, the tools for growth prediction (based on growth kinetics) and real-time growth monitoring by measurement of medium components (based on small-volume analyzing machinery) were installed into the bioreactor system to schedule the operations of medium change and passage and to confirm that culture proceeds as scheduled, respectively. The successive culture of anchorage-dependent cells was conducted with the bioreactor running in an automated way. The automated bioreactor gave a successful culture performance with fair accordance to preset scheduling based on the information in the latest subculture, realizing 79- fold cell expansion for 169 h. In addition, the correlation factor between experimental data and scheduled values through the bioreactor performance was 0.998. It was concluded that the proposed bioreactor with the integration of the prediction and monitoring tools could offer a feasible system for the manufacturing process of cultured tissue products.

[1]  U von Stockar,et al.  On-line determination of animal cell concentration. , 2001, Biotechnology and bioengineering.

[2]  S. Boyce,et al.  Fabrication, quality assurance, and assessment of cultured skin substitutes for treatment of skin wounds , 2004 .

[3]  Michael J Lysaght,et al.  Tissue engineering: the end of the beginning. , 2004, Tissue engineering.

[4]  Robert L Sah,et al.  Perfusion increases cell content and matrix synthesis in chondrocyte three-dimensional cultures. , 2002, Tissue engineering.

[5]  A. Gambhir,et al.  Alteration of cellular metabolism by consecutive fed-batch cultures of mammalian cells. , 1999, Journal of bioscience and bioengineering.

[6]  Masahito Taya,et al.  Evaluation of growth potential of human epithelial cells by motion analysis of pairwise rotation under glucose-limited condition , 2004 .

[7]  H Newman-Gage,et al.  Application of quality assurance practices in processing cells and tissues for transplantation. , 1995, Cell transplantation.

[8]  G. Williams,et al.  Validation of a quality assurance program for autologous cultured chondrocyte implantation. , 1998, Tissue engineering.

[9]  J Winkelman,et al.  Cord lactate, pH, and blood gases from healthy neonates. , 1996, Gynecologic and obstetric investigation.

[10]  F. Hesse,et al.  Developments and improvements in the manufacturing of human therapeutics with mammalian cell cultures. , 2000, Trends in biotechnology.

[11]  M A Levitt,et al.  The effect of the extracellular matrix on the detachment of human endothelial cells , 1984, Journal of cellular physiology.

[12]  Larry V McIntire.  WTEC panel report on tissue engineering. , 2003, Tissue engineering.

[13]  L. Griffith,et al.  Tissue Engineering--Current Challenges and Expanding Opportunities , 2002, Science.

[14]  M Kremer,et al.  Evaluation of dermal-epidermal skin equivalents ('composite-skin') of human keratinocytes in a collagen-glycosaminoglycan matrix(Integra artificial skin). , 2000, British journal of plastic surgery.

[15]  Masahiro Kino-Oka,et al.  Correlation of cellular life span with growth parameters observed in successive cultures of human keratinocytes. , 2002, Journal of bioscience and bioengineering.

[16]  D. Wendt,et al.  The role of bioreactors in tissue engineering. , 2004, Trends in biotechnology.

[17]  J E Prenosil,et al.  Development of an on-line monitoring system of human keratinocyte growth by image analysis and its application to bioreactor culture. , 2000, Biotechnology and bioengineering.

[18]  Kyung A Kang.  Tissue Engineering as a Subdivision of Bioprocess Engineering , 2002, Annals of the New York Academy of Sciences.

[19]  J. Vacanti,et al.  Tissue Engineering and Its Potential Impact on Surgery , 2001, World journal of surgery.

[20]  Anthony Ratcliffe,et al.  Bioreactors and Bioprocessing for Tissue Engineering , 2002, Annals of the New York Academy of Sciences.

[21]  Roland Hetzer,et al.  Tissue-engineering bioreactors: a new combined cell-seeding and perfusion system for vascular tissue engineering. , 2002, Tissue engineering.

[22]  E. M. Dias,et al.  Cell-dislodging methods under serum-free conditions , 1997, Applied Microbiology and Biotechnology.

[23]  M Taya,et al.  Valuation of growth parameters in monolayer keratinocyte cultures based on a two-dimensional cell placement model. , 2000, Journal of bioscience and bioengineering.

[24]  Gary C du Moulin,et al.  Development of a regulatory strategy for the cellular therapies : an American perspective , 2000 .

[25]  K G Vogel,et al.  Effects of hyaluronidase, trypsin, and EDTA on surface composition and topography during detachment of cells in culture. , 1978, Experimental cell research.

[26]  Masahiro Kino-oka,et al.  Growth of Human Keratinocytes on Hydrophilic Film Support and Application to Bioreactor Culture , 1998 .

[27]  Kyriacos A Athanasiou,et al.  Articular cartilage bioreactors and bioprocesses. , 2003, Tissue engineering.