Long-term bone marrow culture-derived stromal fibroblasts as a potential target for gene therapy in acute myelogenous leukemia.
As a part of our continuing efforts to develop gene therapy for acute myelogenous leukemia (AML), this study was undertaken to evaluate the possibility of using autologous bone marrow stromal fibroblasts (BMSFs) as a target cell population. Autologous BMSFs in AML were isolated from the stromal layers of long-term bone marrow culture (LTBMC) using immunomagnetic beads. BMSFs exhibited rapid proliferation even in the absence of growth factors. Cultures stimulated with bFGF produced significantly increased numbers of BMSFs than cultures without added growth factors. Using LNC/LacZ retroviral vector, the transduction efficiency of BMSFs was 13+/-4% at a 5 multiplicity of infection (MOI). LNC/interleukin-2 (IL-2)-transduced BMSFs produced between 1200 and 4800pg of IL-2/10(6) cells per 24h. Using adenoviral vector AdV/LacZ, the transduction efficiency was 84+/-10% at 100, and 92+/-8% at a MOI of 1000. Although the addition of basic fibroblast growth factor, epidermal growth factor, or platelet-derived growth factor did not affect the transduction efficiency, they increased the numbers of transduced cells significantly (P<0.01). AdV/IL-2-treated BMSFs produced high levels of IL-2 over the course of 7 days between 9820 and 22,700pg of IL-2/10(6) cells per 24h. Our finding that the genetically engineered autologous BMSFs of AML could be successfully established in vitro implies that BMSFs obtained from LTBMC might be considered as a target cell population for certain types of clinical gene therapy in AML.
Min
Yoo Hong
YH
Department of Internal Medicine, Yonsei University College of Medicine, Seoul, South Korea. minbrmmd@yumc.yonsei.ac.kr
Li
Guang Xun
GX
Jang
Joon Ho
JH
Suh
Hyung Chan
HC
Kim
Jin Seok
JS
Cheong
June Won
JW
Lee
Seung Tae
ST
Hahn
Jee Sook
JS
Ko
Yun Woong
YW
D000256:Adenoviridae; D001854:Bone Marrow Cells / Q000473:pathology* / Q000502:physiology; D016026:Bone Marrow Transplantation; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D017154:Stromal Cells / Q000473:pathology* / Q000502:physiology; D013997:Time Factors; D014161:Transduction, Genetic; D014182:Transplantation, Autologous; D014407:Tumor Cells, Cultured