Cell Culture
toxicity. The RegeneTOX product line provides tis- sues, customised assays and Contract Testing Services for safety and efficacy assessment of new chemical entities. Included are 2D hepatocytes and 3D liver co-culture from the same liver donor and sibling animal dosing for in vitro 2D to 3D to in vivo comparisons. Skin3 3D full- and partial-thick- ness skin models (based on the Skin2 product line pulled from the market in 1996 and expanded to include immune competent models to address sen- sitisation, inflammation, etc) will be re-launched to address the growing animal alternative market and regulations. Stem cell-derived 3D tissues, including liver, cardiomyocyte and neuronal, for patient-spe- cific and genomic diversity research are in develop- ment. The mission of the company is to provide tis- sues and organs to address medical need regardless of disease progression state; from miniaturised tis- sue for high throughput safety and efficacy assess- ment, to chemical/biological warfare biosensors, molecular diagnostics, personalised medicines, medical devices and tissue implants (Figure 21).
Reinnervate (
www.reinnervate.com) has
researched and developed Alvetex™, a unique polystyrene scaffold designed as a platform tech- nology to enable routine 3D cell culture. The com- pany is now focused heavily on preparing its com- mercial facilities to support quality controlled pro- cedures for the scaled manufacture and production of Alvetex™, due for formal product launch in var- ious formats later this year. Its product quality test- ing levels exceed the various standards required for a consistently high performing cell culture product. This is essential for the global distribution of Alvetex™ to the industrial sector. Its development work includes the manufacture of various devices designed to present the scaffold, to enable optimal function and versatility. For example, well inserts, well clips and large reservoir vessels are all impor- tant ancillary components to maximise the benefits of 3D cell culture using Alvetex™. Reinnervate’s technology is adaptable to scaled manufacture and it has overcome various challenges to deliver a quality 3D cell culture product. While its product is designed as a generic solution to 3D cell culture, scientists at Reinnervate are also developing specif- ic applications for Alvetex™, notably: an artificial 3D skin construct for in vitro testing of cosmetics and topical drugs; a liver toxicity assay using 3D cultured hepatocytes; a 3D platform for enhanced cell differentiation by various stem cell types; and a cell invasion model to evaluate cell migration in 3D to assess various anti-cancer compounds. Alvetex™-enabled 3D cell culture will represent
Drug Discovery World Summer 2010
Figure 22: Reinnervate’s Alvetex™ is supplied as a 200 micron thick membrane that enables cultured cells to form a 3D structure. This sample has been fixed, embedded, transverse sectioned, and cells stained with Toluidine Blue. Alvetex™ is seen in white
the core technology behind each of these applica- tions for which there is significant market demand (Figure 22).
Expansion of hematopoietic stem cells for bone marrow transplantation using conventional 2D culture systems with exogenous growth factors has met with little success. Recent research has shown that the stem cell microenvironment (stem cell niche) plays a key role in the maintenance and expansion of stem cells. Synthecon (
www.synthe-
con.com) has developed, with funding from the National Institutes of Health, a 3D, bio-artificial stem cell niche in its Rotary Cell Culture System (RCCS™) to address this problem. The bio-artifi- cial niche consists of disk-shaped porous scaffolds, bone marrow stromal cells and umbilical cord
Figure 23: Bio-artificial niche assembled in the Rotary Cell Culture System. Bone marrow mesenchymal cells expressing GFP were grown on a porous 3D scaffold. Umbilical cord blood stem cells stained with DilC18(5)-DS were added and allowed to home to the scaffold
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