Friday, September 13, 2013

Three-dimensional cell culturing by magnetic levitation

N3D hits Nature AGAIN! Contact your MIDSCI representative to try this ground breaking 3D cell culture technology. Recently, biomedical research has moved toward cell culture in three dimensions to better recapitulate native cellular environments. This protocol describes one method for 3D culture, the magnetic levitation method (MLM), in which cells bind with a magnetic nanoparticle assembly overnight to render them magnetic. When resuspended in medium, an external magnetic field levitates and concentrates cells at the air-liquid interface, where they aggregate to form larger 3D cultures. The resulting cultures are dense, can synthesize extracellular matrix (ECM) and can be analyzed similarly to the other culture systems using techniques such as immunohistochemical analysis (IHC), western blotting and other biochemical assays. This protocol details the MLM and other associated techniques (cell culture, imaging and IHC) adapted for the MLM. The MLM requires 45 min of working time over 2 d to create 3D cultures that can be cultured in the long term (>7 d). Read more at: http://www.nature.com/nprot/journal/v8/n10/full/nprot.2013.125.html

Tuesday, September 10, 2013

Monoclonal Antibody Production – A discussion on the Culturing of Mouse Hybridoma Cells

The production and maintenance of a hybridoma cell begins with the fusion of a specific antibody producing B cell, to a cancer B cell called a myeloma, which does not produce an antibody by itself. Fusion results in an immortalized line called a hybridoma that will faithfully produce a specific antibody against a single epitope called a monoclonal antibody. Once produced, proper maintenance and culturing is required to maximize the performance and continued production of the antibody in question. These were the topics covered in our last Ask the Expert session – Monoclonal antibody production and the culturing of mouse hybridoma cells. Dr. Fawcett is Director of the BioTechnical Institute of Maryland (BTI) a non-profit institute located in Baltimore, Maryland. He is also the Founder and Director of BioSciConcepts, a social venture of BTI that provides hands-on training for professional scientists in cell culture, baculovirus based expression, as well as topics such as molecular biology, PCR and real-time PCR. Dr. Fawcett who has been in biotechnology for over 30 years, provided real hands-on suggestions that readers could benefit from right away. Question topics included: Use of azaguanine Cell viability post cryopreservation Purity of antigen Serum-free culture Problem obtaining hybridomas in spite of strong signal Culture Conditions Rat-mouse hybridomas Read the answers here: http://cellculturedish.com/2013/07/monoclonal-antibody-production-a-discussion-on-the-culturing-of-mouse-hybridoma-cells/