Establishment of hematological malignancy model in ex vivo cell culture.
10.7534/j.issn.1009-2137.2013.06.002
- Author:
Sheng-Ming ZHAO
1
;
Jian-Xiang WANG
2
;
Hui LIU
3
;
Ming-Ting PENG
4
Author Information
1. 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.
2. Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
3. Department of Hematology,Beijing Hospital, Beijing 100730, China.
4. National Center for Clinical Laboratory, Ministry of Public Health of China.
- Publication Type:Journal Article
- MeSH:
Animals;
Bone Marrow Cells;
pathology;
Cell Culture Techniques;
Genetic Vectors;
Hematologic Neoplasms;
Humans;
Mice;
Mice, Inbred C57BL;
Neoplasms, Experimental;
Oncogene Proteins, Fusion;
genetics;
Retroviridae;
genetics
- From:
Journal of Experimental Hematology
2013;21(6):1373-1379
- CountryChina
- Language:Chinese
-
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.
