AVS 53rd International Symposium
    Biomaterial Interfaces Tuesday Sessions
       Session BI-TuA

Paper BI-TuA2
Compartmentalized Bioreactor: In Vitro Model for Osteogenesis and Breast Cancer Metastasis

Tuesday, November 14, 2006, 2:20 pm, Room 2001

Session: Cells at Surfaces
Presenter: D. Ravi, Pennsylvania State University
Authors: D. Ravi, Pennsylvania State University
L.A. Shuman, Pennsylvania State University
A.M. Mastro, Pennsylvania State University
C.V. Gay, Pennsylvania State University
E.A. Vogler, Pennsylvania State University
Correspondent: Click to Email
An advanced bioreactor that permits long-term study (up to 10 months) of cell/protein mediated processes such as bone accretion and cancer metastasis was developed and tested. The bioreactor based on the principle of simultaneous-cell-growth-and-dialysis, separates a cell growth chamber from a media reservoir by a dialysis membrane, compartmentalizing cell growth and cell nutrition functions. Extraordinarily stable culture conditions afforded by the reactor sustained mouse calvarial osteoblasts (MC3T3-E1, ATCC CRL-2593) for periods up to 10 months without the need for sub-culture. Months-long culture resulted in the formation of macroscopic (mm scale) sheets of bone over the inner surface of the dialysis membrane. Development of three-dimensional, tissue-like biosynthetic bone in the reactor was followed by light microscopy, scanning electron and transmission electron microscopy. Markers of osteoblast differentiation (alkaline phosphatase) and mineralization (Von Kossa Assay, SEM-EDS, XRD) were used to analyze the progression of isolated osteoblast inoculum to highly collagenous mineralizing tissue. In the second part of the study, the biosynthetic bone tissue was challenged by co-culture with GFP-expressing metastatic breast cancer cells (MDA-MB-231). Effect of the cancer cells on the morphology and organization of bone tissue was followed by confocal, scanning and transmission electron microscopy. Co-culture with breast cancer cells resulted in migration of cancer cells through the osteoblast tissue, disruption of the collagenous matrix, apoptosis and increased production of inflammatory cytokines (IL-2, IL-6). Compartmentalized bioreactor permits development of mineralizing 3-D collagenous bone tissue from isolated osteoblast inoculum over extended time periods up to 10 months and is an ideal in vitro vehicle for studying long-term cell-cell interactions involved in osteogenesis and osteopathology that are inaccessible to conventional cell culture techniques.