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The FMCA-GM assays, high throughput non-clonogenic alternatives to CFU-GM in preclinical hematotoxicity testing
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- Haglund, Caroline (author)
- Uppsala universitet,Klinisk farmakologi,Cancer Pharmacology and Informatics/Rolf Larsson
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- Åleskog, Anna (author)
- Uppsala universitet,Klinisk farmakologi,Cancer Pharmacology and Informatics/Rolf Larsson
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- Håkansson, Lena Douhan (author)
- Uppsala universitet,Institutionen för medicinska vetenskaper,Division of Haematology
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- Jacobsson, Stefan (author)
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- (creator_code:org_t)
- Elsevier BV, 2010
- 2010
- English.
- In: Toxicology Letters. - : Elsevier BV. - 0378-4274 .- 1879-3169. ; 194:3, s. 102-107
- Journal article (peer-reviewed)
Abstract
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- One of the most common dose limiting adverse effects in cancer treatment is myelotoxicity. The aim of this study was to develop an in vitro method for measuring potential myelotoxic properties of a drug candidate in a high throughput setting. Human CD34+ progenitor cells from umbilical cord blood were plated in 384-well microplates with drugs in liquid culture, supplemented with specific cytokines for the granulocytopoietic-macrophage lineage. After 7 or 14 days of proliferation and differentiation the cells were analyzed using the automated non-clonogenic fluorometric microculture cytotoxicity assay (FMCA). Two types of assays setups were evaluated, the FMCA-GM7 where cells were exposed to drugs directly after thawing and cytotoxicity measured on day 7 in contrast to the FMCA-GM14 where the cells were cultured 7 days prior to plating and drug exposure, with viability analysis on day 14 of differentiation. Drug sensitivity was similar in both assays and method validation was performed using 24 drugs with known myelotoxic profile (acyclovir, bortezomib, busulfan, carboplatin, chloramphenicol, chlorpromazine, cisplatin, cytarabine, clozapine, doxorubicin, erlotinib, etoposide, 5-fluorouracil, fludarabine, gefitinib, gemcitabine, hydroxyurea, imatinib, lomustine, melphalan, sorafenib, sunitinib, taxol and 6-thioguanine). The 50% inhibitory concentrations (IC50) from the FMCA-GM7 and the FMCA-GM14 correlated highly (r = 0.83) and (r = 0.82), respectively, with IC50 from the established clonogenic assay (CFU-GM), obtained from the literature. The current data suggests that the FMCA-GM could offer a simple and robust alternative to the CFU-GM assay in preclinical hematotoxicity studies.
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