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A novel method for the generation of reaggregated organotypic cultures that permits juxtaposition of defined cell populations
Author(s) -
Sheridan Julie M.,
Taoudi Samir,
Medvinsky Alexander,
Blackburn C. Clare
Publication year - 2009
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20505
Subject(s) - microbiology and biotechnology , biology , haematopoiesis , mesonephros , in vitro , cell , stem cell , biochemistry , embryonic stem cell , gene
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. genesis 47:346–351, 2009. © 2009 Wiley‐Liss, Inc.