title: Effects of radiation and 4-hydroperoxycyclophosphamide on production of G- and GM-CSF by stromal cells.
abstract: The effects of in vitro radiation and exposure to 4-hydroperoxycyclophosphamide (4-HC) on the production of G- and GM-CSF by different components of the human hematopoietic microenvironment are described. The marrow microenvironment is composed of fibroblasts, endothelial cells, macrophages, and adipocytes. To study the effects of radiation/4-HC on colony-stimulating factor (CSF) production by stromal cells, confluent layers of umbilical cord endothelial cells (EC), marrow fibroblasts (MF), and heterogeneous adherent layers (HAL) derived from long-term marrow cultures were established. These layers were exposed to radiation up to 3000 cGy and/or 100 mumol/ml of 4-HC and subsequently stimulated with IL-1 beta on day 0, 7, or 14 after radiation/4-HC. Following IL-1 exposure conditioned medium (CM) was collected and G- and GM-CSF levels were measured by ELISA and their ability to support colony formation was assessed. G- and GM-CSF levels after exposure to 4-HC and radiation were 12,460 +/- 172 pg/ml and 2268 +/- 160 pg/ml for EC, 2214 +/- 94 pg/ml and 263 +/- pg/ml for MF, and 3168 +/- 316 pg/ml and 356 +/- 34 pg/ml for HALs, respectively. For each cell group there was no significant difference between levels obtained without exposure and levels after exposure to 4-HC and/or radiation (p > 0.6). Comparison of levels obtained from different cell groups showed significant differences with EC media being the highest (p < 0.0001). To test the activity of these measured factors, CM of different sources was used in colony assays of CD 34+ cord blood progenitors.(ABSTRACT TRUNCATED AT 250 WORDS)
mesh_terms: D001853:Bone Marrow / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D016026:Bone Marrow Transplantation; D003114:Colony-Forming Units Assay; D017077:Culture Media, Conditioned; D003520:Cyclophosphamide / Q000031:analogs & derivatives / Q000494:pharmacology; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis*; D006801:Humans; D066298:In Vitro Techniques
chemical_list: D017077:Culture Media, Conditioned; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D003520:Cyclophosphamide; C011272:perfosfamide