title: [Establishment of hematological malignancy model in ex vivo cell culture].
abstract: The aim of this study was to develop an ex vivo cell culture system for establishing the hematological malignancy model. Mouse bone marrow cells were transfected with GFP-expressed retroviral vectors encoding various leukemia/lymphoma-associated fusion proteins (TEL-PDGFR, Rabaptin5-PDGFR, p210BCR-ABL, AML1-ETO, NPM-ALK). After transfection, the cells were cultured in IMDM containing 10% FCS without growth factors, or with one of the following growth factor combinations: (1) murine c-kit ligand (KL) plus human flt3 ligand (FL); (2) IL-3, thrombopoietin, G-CSF, and hyper-IL-6 (3/T/G/H6); (3) KL/FL plus 3/T/G/H6. The flow cytometry was used to detect the ability of combinations of growth factors to complement the oncogene fusion protein to support self-renewal of the transfected cells. The results showed that the transfected cells could be amplified sustainably in the logarithmic growth way. The indicated combination of c-Kit ligand (KL) with flt-3 ligand (FL) supported the self-renewal of the marrow cells transfected with vectors encoding TEL-PDGFR, Rabaptin5-PDGFR, AML1-ETO and NPM-ALK, in addition to KL/FL, the self-renewal of p210 BCR-ABL transfected-marrow cells also required IL-3. The morphology of cells emerged from culture can be the predictor of the corresponding oncogene-associated malignancy. It is concluded that this study establishes a culture system ex vivo which provides a generalized method for studying hematological malignancies, and may facilitate the screening for therapeutic agents.
authors.affiliation: Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences(CAMS), Beijing 100730, China; Department of Hematology,Beijing Hospital, Beijing 100730, China. E-mail:shengmingzhao@sina.com.
authors.forename: Sheng-Ming
authors.initials: SM
authors.lastname: Zhao
authors.affiliation: Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
authors.forename: Jian-Xiang
authors.initials: JX
authors.lastname: Wang
authors.affiliation: Department of Hematology,Beijing Hospital, Beijing 100730, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: National Center for Clinical Laboratory, Ministry of Public Health of China.
authors.forename: Ming-Ting
authors.initials: MT
authors.lastname: Peng
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000473:pathology*; D018929:Cell Culture Techniques; D005822:Genetic Vectors; D019337:Hematologic Neoplasms; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009374:Neoplasms, Experimental; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D012190:Retroviridae / Q000235:genetics
chemical_list: D015514:Oncogene Proteins, Fusion