Frye Cynthia A, Patrick Charles W
Laboratory of Reparative Biology and Bioengineering, Department of Plastic Surgery, University of Texas M. D. Anderson Cancer Center, Houston, TX 77230-1402, USA.
In Vitro Cell Dev Biol Anim. 2006 May-Jun;42(5-6):109-14. doi: 10.1290/0509055.1.
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.
本文介绍了为培养和分析三维（3-D）脂肪样组织而开发的技术，以此作为一种手段，弥合当前前脂肪细胞（PA）培养的局限性与提供临床上有用的脂肪组织体积之间的差距，这些脂肪组织体积对重建手术中的软组织工程策略具有重要意义。进行了初步研究，以确定可视化和分析三维PA样组织的技术，并制定在有和没有微载体的情况下在高纵横比容器旋转壁生物反应器中培养3T3-L1细胞的成功策略。接下来，进行了一系列培养实验，以验证这些技术，并比较有和没有微载体时细胞的培养情况。最后，使用灌注旋转壁生物反应器进一步研究所生成的聚集体或组织的性质。通过组织学和体内动物研究分析了在灌注系统中形成的聚集体。成功获得了直径达4-5毫米、无微载体、能够进行脂质加载且由活细胞组成的PA样组织。我们已成功证明，大型组织聚集体可在生物反应器培养系统中生长。
