Immunofluorescence Visualization of Fibroblast or Cancer Cells in RAFT™ 3D Cell Cultures

3D culture is transforming cell biology research and tissue engineering applications. These advanced tools are allowing researchers to develop higher-order structures with cells in vitro. This, in turn, allows cells to grow and interact in an environment more closely mimicking in vivo. However, such revolution in research is also coupled to new challenges. It can be very difficult to apply standard cell analysis techniques on 3D cultures which have long been established in 2D environment. The tight structure of some spheroids, which form a necrotic core, the lack of transparency of many plastic materials used in 3D methods or the dense fiber network of some 3D hydrogels can interfere with basic techniques such as imaging, transfection, cytotoxicity assays or biotherapeutic applications.

The RAFT™ 3D Cell Culture System is attempting to address such challenges with 3D methods enabling researchers to work with a 3D system that is well supported with protocols to conduct downstream assays. Lonza continues to develop and support the RAFT™ System with additional optimized protocols that allow for applying standard histological, biochemical and imaging techniques to 3D cultures.

This white paper shows how the breast cancer cell line MCF7 and human dermal fibroblasts were cultured using the RAFT™ 3D System for several days prior to fixation and immunocytochemical staining. The white paper demonstrates an efficient permeability of the antibodies through the RAFT™ Matrix to visualize these 3D cultures.

Find more RAFT™ 3D Cell Culture White Papers at www.lonza.com/raft-applications.