微组织使用胶原蛋白的模块的制备凝胶含有细胞
Summary
使用圆柱胶原凝胶，称为模块，其中包含嵌入细胞表面的微组织的创建是涂有内皮细胞。
Abstract
这个协议描述的称为模块的微观组织类型制造。该模块的方法生成统一的，可扩展性和吻合血管的组织。该模块可制成胶原蛋白，以及其他gelable或交联材料。他们后制造的直径约2毫米长，0.7毫米，但在尺寸缩小与嵌入式细胞或模块与内皮细胞涂。单独的模块都足够小，嵌入细胞内的氧气和其他营养物质的扩散限制，但模块可以打包在一起，形成较大的组织，perfusable。因为不同类型的细胞，可以嵌入在或涂在模块之前，他们挤在一起，形成复杂的组织，这些组织都采用模块化施工。模块有三个主要步骤：（1）中的胶原蛋白和嵌入在它的细胞，（2）凝胶在管内的胶原蛋白和切割与内皮细胞的模块和（3）涂层的模块。
Protocol
1）准备油管切割2至3米长度的聚乙烯管（0.76毫米内径× 1.22毫米外径）。 20 G1的针管端成一个主题。 线圈油管并将其放置在一个转换器，自密封袋（7 1 / 2“× 13”）。油管的两端应置于袋启封，并在地方与气体消毒磁带录音。 密封袋和气体消毒。 2）中和胶原蛋白注：1毫升的胶原蛋白填充聚乙烯管材约2米。 将所…
Discussion
我们编造了几个不同的微观组织，使用与不同的细胞类型1-11嵌入式模块。我们已经成功地嵌入原发性心肌细胞，胰岛，脂肪干细胞和间质基质细胞以及一些包括肝癌，NIH 3T3和克隆肝细胞的细胞株。我们涂有几种类型包括大鼠主动脉血管内皮细胞，人脐静脉内皮细胞和人类微血管内皮细胞内皮细胞的模块。模块也已产生胶原蛋白和poloxamine聚合物或含有药物洗脱微球（图4）的混合物。 ?…
Declarações
The authors have nothing to disclose.
Acknowledgements
经费是由美国国立卫生研究院（EB 001013），自然科学和工程研究理事会，加拿大和加拿大卫生研究院。我们感谢美联社麦圭根博士和她的专长，帮助模块的开发。
Materials
| Material Name | Tipo | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Intramedic tubing PE60 | Becton Dickson | 427416 | Different diameter of tubing can be used to change the diameter of the modules | |
| Phosphate-buffered saline (PBS) | Gibco | 20012-027 | ||
| Trypsin-EDTA | Gibco | 25200-072 | ||
| Purcol acidificed collagen, 3 mg/mL | Cedarlane | 5005-B | ||
| Sodium bicarbonate | Sigma-Aldrich | S5761 | ||
| 20G needle | Becton Dickson | 305175 | Diameter of the needle needs to be similar to the diameter of the tubing | |
| 3 mL syringe | BD Biosciences | 309585 | ||
| 15 mL tube | BD Biosciences | 352096 | ||
| 50 mL tube | BD Biosciences | 352070 | ||
| Convertors Self-Seal Pouch 7 1/2” x 13” | Cardinal Health | 92713 | ||
| 10 ml Wide Tip serological pipet | BD Falcon | 357504 | ||
| 10 ml serological pipet | BD Falcon | 357551 | ||
| 5 ml serological pipet | BD Falcon | 357543 |
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Citar este artigo
Chamberlain, M. D., Butler, M. J., Ciucurel, E. C., Fitzpatrick, L. E., Khan, O. F., Leung, B. M., Lo, C., Patel, R., Velchinskaya, A., Voice, D. N., Sefton, M. V. Fabrication of Micro-tissues using Modules of Collagen Gel Containing Cells. J. Vis. Exp. (46), e2177, doi:10.3791/2177 (2010).