Cell Culture
ly on the need for isolation efficiency and prolifera- tion, without consideration of these parallel require- ments of a 3D model. In combination with primary human keratinocytes isolated in CELLnTEC’s Progenitor Cell Targeted media, this 3D medium has been found to establish 3D epidermal models with accurate representation of the in vivo structure, marker expression and lipid profile and to maintain them for an extended period. CELLnTEC also has a thriving CRO business testing compounds on these skin models. Additionally, outside laboratories are using its cells and media for 3D Airway models, corneal transplantation research and 3D co-culture experiments. CELLnTEC has recently invested in new facilities and staff to extend its offering of in vitro 3D models both via CRO and packaged prod- ucts (Figure 12).
Figure 12: A fully differentiated 3D epithelial model, established from a population of epidermal keratinocyte progenitor cells isolate in CELLnTEC Progenitor Cell Targeted medium and differentiated in CELLnTEC optimised 3D medium
microlitres/day. An innovative microfabricated per- fusion barrier promotes uniform nutrient transport into the gel chamber. They key advantages of the 3D:M design are: 1) enabling 3D perfusion screen- ing; 2) high quality cell imaging in a 3D environ- ment; and 3) reduction of cell/gel usage by 10 times. CellASIC has also developed variations of the 3D microfluidic culture concept to address applications requiring: larger gel chambers, automated solution exchange, maintaining long term spatial gradients, and in-chamber immunostaining (Figure 11).
CELLnTEC Advanced Cell Systems (www.cell-
ntec.com) applies the latest developments in adult stem cell signalling to formulate speciality cell cul- ture media unique to the epithelial market. CELLnTEC began as a CRO developing 3D vaginal and bladder models. This led to a number of epithe- lial culture media, and primary cells isolated in these media especially suitable to 3D cell culture. CELLnTEC has optimised a skin medium that sup- ports the conflicting requirements of 3D in vitro modelling. 3D models place very specific demands on the cell culture medium. Specifically, the medium must encourage cells to reach terminal differentia- tion and stratify, while in parallel maintaining the population of proliferative cells necessary in the basal layer to establish a fully stratified model. These conflicting requirements are poorly addressed by conventional media, which were developed sole-
32
The ‘Cell Cycle’ is a partnership formed between the European Collection of Cell Cultures (ECACC) (
www.hpacultures.org.uk), Sigma Aldrich (
www.sigmaaldrich.com), Corning (
www.corn-
ing.com) and XCellR8 (
www.x-cellr8.com). This partnership fully recognises that proliferation and differentiation of cells in culture has become increasingly important for basic research, drug dis- covery, tissue engineering and regenerative medi- cine. The way in which cells proliferate and differ- entiate is defined by the unique microenvironments required by the different cell types in a multicellu- lar organism. Several factors influence the cells’ proliferative and differentiation status and these fall into two broad categories: soluble cues (growth factors, metabolites, dissolved gases) and insolu- ble, physical cues (the composition, architecture,
Figure 13: Human mesenchymal stem cells grown by ECACC for five days on the surface of a HyStem hydrogel with non-covalently incorporated collagen I
Drug Discovery World Summer 2010
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