|Title||A novel method for the generation of reaggregated organotypic cultures that permits juxtaposition of defined cell populations.|
|Publication Type||Journal Article|
|Year of Publication||2009|
|Authors||Sheridan JM, Taoudi S, Medvinsky AL, C Blackburn C|
|Date Published||2009 May|
|Keywords||Animals, Cell Aggregation, Cell Lineage, Cells, Cultured, Embryo, Mammalian, Female, Flow Cytometry, Fluorescent Antibody Technique, Green Fluorescent Proteins, Immunohistochemistry, Keratins, Male, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Transgenic, Organ Culture Techniques, Pregnancy, Thymus Gland|
Cellular reaggregation methods have been used to generate in vitro organotypic cultures as a means to elucidate the cellular and molecular requirements of organogenesis. However, reproducibility from experiment to experiment has remained problematic and furthermore, current protocols do not support reaggregation of many important tissues. Here, using the thymus as a model organ, we present a novel reaggregation method termed "compaction reaggregation" that offers improved kinetics of reaggregation and greatly improved efficiency. Using compaction reaggregation we have been able to reaggregate the aorta-gonad- mesonephros region, a tissue that previously proved refractory to commonly used reaggregation methods, enabling the study of hematopoietic stem cell emergence and expansion. Additionally, compaction reaggregation permits the juxtaposition of different cell layers within the aggregated structure thus providing the means to study inductive interactions between different cell populations in vitro.
|Grant List||G0500950 / / Medical Research Council / United Kingdom|