Three-dimensional adipose tissue model using low shear bioreactors.
Presented here are techniques developed to culture and analyze three-dimensional (3-D) adipose-like tissues as a means to bridge the gap between current limitations in culturing preadipocytes (PAs) and that of providing clinically relevant volumes of adipose tissue useful for soft tissue engineering strategies in reconstructive surgery. Pilot studies were performed to determine techniques to visualize and analyze 3-D PA-like tissues as well as to develop successful strategies to culture 3T3-L1 cells in a high aspect ratio vessel rotating-wall bioreactor both with and without microcarriers. Next, a series of cultures were accessed to verify these techniques as well as to compare the culture of the cells with and without microcarriers. Finally, a perfused rotating-wall bioreactor was used to further investigate the nature of the aggregates or tissues being generated. The aggregates that formed in the perfused system were analyzed via histology and in vivo animal studies. PA-like tissues as large as 4-5 mm in diameter without microcarriers that were capable of lipid-loading and composed of viable cells were achieved. We have successfully demonstrated that large tissue aggregates can be grown in bioreactor culture systems.
Frye
Cynthia A
CA
Laboratory of Reparative Biology and Bioengineering, Department of Plastic Surgery, University of Texas M. D. Anderson Cancer Center, Houston, TX 77230-1402, USA.
Patrick
Charles W
CW
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology*; D000273:Adipose Tissue / Q000166:cytology* / Q000254:growth & development; D000818:Animals; D019149:Bioreactors; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D020099:Coated Materials, Biocompatible; D003094:Collagen / Q000737:chemistry; D051379:Mice; D008863:Microspheres; D010865:Pilot Projects; D051381:Rats; D046509:Tissue Culture Techniques / Q000295:instrumentation; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods
D020099:Coated Materials, Biocompatible; D003094:Collagen