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The role of recombinant haematopoietic growth factors in human long‐term bone marrow culture in serum‐free medium
Author(s) -
Douay Luc,
Giarratana MarieCatherine,
Drouet Xavier,
Bardinet Dominique,
Gorin NorbertClaude
Publication year - 1991
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1991.tb08002.x
Subject(s) - haematopoiesis , erythropoietin , granulopoiesis , bone marrow , granulocyte macrophage colony stimulating factor , granulocyte , erythropoiesis , progenitor cell , immunology , colony stimulating factor , andrology , cfu gm , biology , stromal cell , stem cell , medicine , endocrinology , cytokine , microbiology and biotechnology , cancer research , anemia
We have previously reported prolonged in vitro maintenance of human bone marrow progenitor cells using a serum‐free (SF) liquid culture system. The present study was undertaken to determine recombinant growth factor (rGF) requirement of long‐term marrow culture (LTMC) in absence of exogenous serum, to avoid interference of any undefined components. Our data clearly show that the presence of serum is a major obstacle to the correct evaluation of rGF activity. However, in SF conditions the sequential analysis of these rGFs, alone or in combination, clearly showed a stimulating activity of interleukin 3 (IL3), granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and erythropoietin (EPO) on granulopoiesis and erythropoiesis. Indeed, the cumulative number of progenitors recovered during an 8‐week period exceeded the initial input by a fractor of 1·7 for granulocyte‐macrophage (CFU‐GM), of 3 for erythroid blast‐forming units (BFU‐E) and of 5·45 for CFU‐E when EPO, GM‐CSF and IL3 were combined. This study has confirmed that the system is able to sustain haematopoiesis for 8 weeks in a way similar to that in serum‐dependant LTMC, despite diminished stromal adherent layer development which never covered more than 50% of the flask surface. We conclude that this defined SF‐LTMC system provides a reproducible technique for studying human haematopoiesis.