Cell Culture
precise microscopic analysis and excellent cell number-size correlation; and d) the possibility to increase the complexity from homotypic to het- erotypic tissue models. These tissues are provided off-the-shelf or custom-made in 96-well-compati- ble plates to allow for a straightforward upgrade from 2D to 3D at the user lab using existing instruments and assay procedures (Figure 17).
Kuraray (
www.kuraray.co.jp) is developing a new cell culture plate called the Micro-Space Cell Culture plate by utilising its micro-fabrication technology. The plate has a number of micrometer size compartments regularly arrayed on its surface which provide cells ‘micro-space’ to form 3D struc- ture. The plate has several advantageous features: it conforms to the standard microplate footprint enabling simple handling; it has good observabili- ty; and there is uniformity in the size or shape of the microstructure. No special techniques, includ- ing gel formation, are required. The bottom of the plate is a thin film made of transparent material, polystyrene, so that it is suitable for microscopic observation for fluorescent immunoassay. The size of the cell aggregates can be controlled by the size of the micro-pattern used. The shape of the micro- pattern can be designed as desired. It is believed that an optimum micro-pattern varies depending on cell type. For instance, primary hepatocytes form a spheroid-like structure in a square com- partment (eg surface which width x 100µ depth) (see Figure 18). Hepatocyte-specific functions, for example albumin secretion and some of cytochrome P450s activity, were well preserved on the Micro-Space Cell Culture plate compared to those on a conventional flat plate by several-fold as measured by mRNA expression. On the other hand, cardiomyocytes form cell networking on another pattern providing synchronous contrac- tion, and mammary epithelial cells present acinus- like structure on other pattern showing similar morphology to that seen in vivo. The Micro-Space Cell Culture plate is currently nearing commercial- isation. Kuraray has prepared 24-well and 96-well sample plates for collaborators in Academia and Pharma, but other formats 6-, 12- and 384-well will be made if Kuraray confirms demand. Kuraray believes the plate is applicable for HTS due to the simple handling and good observable feature.
QGel (
www.qgelbio.com) recently commercialised QGel™ MT 3D Matrix (QGel™), a synthetic hydrogel supported by more than 10 years of sci- entific experimentation and publications (Figure 19). Among many of its applications, QGel™ can
Drug Discovery World Summer 2010
Figure 18: Microscopic observation of primary hepatocytes on Kuraray Micro-Space Cell Culture plate
be used as a matrix for 3D cell culture and is par- ticularly interesting for studies in regenerative medicine, cancer research and drug screening. The synthetic biological and biochemical components used to bioengineer QGel™ extracellular matrix (ECM) are what make QGel™ unique compared to currently available products. Protease-sensitive sites and cell-adhesion ligands, such as RGD (pep- tide containing the amino acid sequence Arginine- Glycine-Aspartic Acid), are key components of QGel™ that mimic essential features of natural ECM (Figure 19). Unlike other synthetic products, QGel™ degrades like natural ECM, ie by cell- secreted and activated proteases, such as MMP (matrix metalloproteinase). QGel™ components
Figure 19: QGel™ is synthetic gel that consists of a polymer (a) structural component, (b) protease sensitive sites, and (c) cell- adhesion ligands. These three components are pre-mixed as a powder in a single vial. After solving the powder with QGel™ Buffer (not shown) a (d) 3D hydrogel structure forms that mimics key features of the natural extracellular matrix. Gelation occurs in 5- 10 minutes, which allows you to encapsulate cells and shape the gel as you wish, such as in the form of a (e) 3D disc. QGel™ matrix products are available as (f) non-degradable and (g) degradable gels (with and without incorporated RGD-peptide) delivered sterile in ready-to-use vials for 0.5mL of gelated matrix
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