The liquid overlay technique is the key to formation of co-culture spheroids consisting of primary osteoblasts, fibroblasts and endothelial cells
The liquid overlay technique is the key to formation of co-culture spheroids consisting of primary osteoblasts, fibroblasts and endothelial cells
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DOI：
10.3109/14653249.2011.583233
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发表时间：
2011-09-01
期刊：
影响因子：
4.5
通讯作者：
Oberringer, Martin
中科院分区：
文献类型：
--
作者：
Metzger, Wolfgang;Sossong, Daniela;Oberringer, Martin
背景目标。各种细胞类型的三维（3-D）培养比二维（2-D）细胞培养技术更精确地反映体内情况。球状体作为三维细胞构建体已经在肿瘤研究中使用了很长时间。它们也被用于研究血管生成机制，这对于许多组织工程方法的成功至关重要。已知几种形成球状体的方法，但是缺乏评价这些技术的性能的系统研究。方法.我们评价了悬滴技术、羧甲基纤维素技术和液体覆盖技术形成由原代成骨细胞、成纤维细胞和内皮细胞组成的单一和共培养球体的性能。这三种技术的性能进行了评估的产率和重现性。系统地确定了生成的球状体的大小。结果液体覆盖技术是最适合于可重复地产生球状体的技术。该技术的产率为60%和100%之间的单培养球状体和100%的共培养球状体。通过改变一个球状体中组织的接种细胞的数量，可以容易且精确地调节球状体的大小。由三种不同细胞类型组成的共培养球体的形成是可能的。结论.我们的研究结果表明，最合适的技术形成球状体可以从所选择的细胞类型，特别是如果使用原代细胞。由三种不同类型的细胞组成的共培养球体将在进一步的研究中用于研究血管生成现象。
Background aims. The 3-dimensional (3-D) culture of various cell types reflects the in vivo situation more precisely than 2-dimensional (2-D) cell culture techniques. Spheroids as 3-D cell constructs have been used in tumor research for a long time. They have also been used to study angiogenic mechanisms, which are essential for the success of many tissue-engineering approaches. Several methods of forming spheroids are known, but there is a lack of systematic studies evaluating the performance of these techniques. Methods. We evaluated the performance of the hanging drop technique, carboxymethyl cellulose technique and liquid overlay technique to form both mono-and co-culture spheroids consisting of primary osteoblasts, fibroblasts and endothelial cells. The performance of the three techniques was evaluated in terms of rate of yield and reproducibility. The size of the generated spheroids was determined systematically. Results. The liquid overlay technique was the most suitable for generating spheroids reproducibly. The rate of yield for this technique was between 60% and 100% for monoculture spheroids and 100% for co-culture spheroids. The size of the spheroids could be adjusted easily and precisely by varying the number of seeded cells organized in one spheroid. The formation of co-culture spheroids consisting of three different cell types was possible. Conclusions. Our results show that the most suitable technique for forming spheroids can vary from the chosen cell type, especially if primary cells are used. Co-culture spheroids consisting of three different cell types will be used to study angiogenic phenomena in further studies.