eCM (Eur Cell Mater / e Cells & Materials) Not-for-profit Open Access
Created by Scientists, for Scientists
 ISSN:1473-2262         NLM:100973416 (link)         DOI:10.22203/eCM

2010   Volume No 19 – pages 166-179

Title: Novel in vitro co-culture methodology to investigate heterotypic cell-cell interactions

Author: EF Burguera, M Bitar, A Bruinink

Address:Osteoarticular and Aging Research Lab., Biomedical Research Center – INIBIC
Complejo Hospitalario Universitario A Coruña, 15006 A Coruña, Spain

E-mail: elena.fernandez.burguera at

Key Words: Biomaterial surfaces, 2D-co-culture, osteoblasts, cell proliferation, fibroblasts, in vitro.

Publication date: April 23rd 2010

Abstract: Cell-cell interactions are of crucial importance for the formation of tissues, homeostasis and regeneration processes as well as reactions on foreign bodies including implants. So far, however, the importance of heterotypic cell-cell interactions in the in vitro evaluation of implant surfaces has been largely neglected. This work aims to develop an in vitro methodology that enables the in-depth investigation of heterotypic cell-cell interactions in a mixed co-culture system, and to validate it with a primary adult human bone-derived osteoblast cells (HBCs) – abdominal fibroblasts (HAFs) system. The methodology proposed combines a simple live labelling step, semiautomated fluorescence image acquisition and analysis to characterize the interactions between different cell types (cell population dynamics) in co-culture in terms of cell proliferation and cell spatial distribution of each cell type. In this co-culture system, direct cell-cell contacts between the two cell types were permitted while the determination of cell-type specific responses could still be elucidated. We could show that HAF proliferation was reduced in a way negatively correlated with the seeding HBC/HAF ratio, i.e., a high proportion of HBC in the co-culture had an inhibitory effect on HAF proliferation. In all cultures segregation was found after 4 and 7 days of co-culture. HBCs were segregated at low ratios while HAFs were segregated at high ratios. Cell-cell distances depended on the total cell number in the co-culture but the dependence was different for each cell type.

Article download: Pages 166-179 (PDF file)
DOI: 10.22203/eCM.v019a17