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Modification of human long‐term bone marrow cultures: Establishment of a functional stromal microenvironment devoid of myeloid progenitors
Author(s) -
Moreau Isabelle,
Andreoni Christine,
Caux Christophe,
Saeland Sem,
Rigal Dominique
Publication year - 1992
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.1992.tb00910.x
Subject(s) - stromal cell , haematopoiesis , bone marrow , cd34 , progenitor cell , myeloid , myelopoiesis , clonogenic assay , biology , microbiology and biotechnology , flow cytometry , immunology , stem cell , cancer research , cell culture , chemistry , pathology , medicine , genetics
Differences in the plastic adhesive properties of bone marrow (BM) cells were used to initiate modified stromal layers (MSL) from long‐term cultures by removing non‐adherent cells shortly (4 to 18 hours) after initial seeding. Following this early modification, adherent cells generated a confluent layer after 21 days of culture. Cellular characteristics of volume and spontaneous fluorescence determined by flow cytometry showed that the MSL included 82% fibroblastic stromal cells, 8% macrophages and 10% myelomonocytic cells. Furthermore, clonogenic assays revealed that the MSL were devoid of hematopoietic progenitor cells. MSL were found to sustain long‐term myelopoiesis for at least 7 weeks from exogenously added hematopoietic progenitors isolated from bone marrow (CD34+ cells), thereby demonstrating their functionality. The present experimental model appears of interest for the study of interactions between defined populations of hematopoietic cells and cells of the adherent layer. Of importance, our present modifications of human long‐term bone marrow culture are technically simple and do not involve manipulation of the stromal cells.