Malignant transformation of host stromal fibroblasts derived from the bone marrow traced in a dual-color fluorescence xenograft tumor model

Abstract

Abstract. Solid tumors are abnormal tissues containing tumor and non-tumor cells, also known as tumor stromal cells. However, the malignant potential of tumor stromal cells remains largely unknown. The aim of the present study was to investigate the malignant potential of host bone marrow-derived stroma cells in transplanted subcutaneous tumors of the glioma stem/progenitor cells (GSPCs) labeled using the dual-color fluorescent tracer technique. The previously established human glioma stem/progenitor cell line SU3 was transfected with red fluorescence protein (SU3-RFP) and transplanted subcutaneously into green fluorescent protein (GFP) transgenic nude mice and chimeric mice in which GFP was only expressed by bone marrow-derived cells (BMDCs). The xenograft tumors were subcultured in vitro and two immortalized GFP-expressing stromal cell lines were cloned from the transplanted tumors. The two cloned cell lines showed an accelerated growth rate, loss of cell contact inhibition, high cloning efficiency, and high DNA content and telocentric (murine) chromosomes with heteroploid characteristics. The tumorigenesis rate (10/10, 1x10 6 ) of these host stromal cells was further evidence of malignant transformation. Immunofluorescence assay of the two host cell lines showed that they expressed fibroblast markers such as FAP, S100A4 and α-SMA, as well as mesenchymal cell markers such as CD44 and CD105. In conclusion, bone marrow-derived stromal fibroblasts recruited to tumors have the potential for malignant transformation induced by the tumor microenvironment, which provides new evidence for the role of the stroma in malignant transformation.

Novel enhanced GFP‐positive congenic inbred strain establishment and application of tumor‐bearing nude mouse model

“…With a fluorescent tracer xenograft model based on the inbred nude mice that express EGFP extensively, including in brain tissue, our work focused on TME research has confirmed that some host mesenchymal cells have undergone malignant transformation. In the present as well as our previous reports, 12,20,21 we found that some fibroblasts, bone marrow‐derived dendritic cells and oligodendrocytes in transplanted tumor microenvironmentwere malignant transformation induced by primary transplanted tumor cells. The Foxn1 nu .B6‐Tg(CAG‐EGFP) mice auricle transplantation model could also facilitate research on the tumor immune microenvironment in vivo, with a two‐photon fluorescence microscope.…”

Coaxial 3D bioprinting of self-assembled multicellular heterogeneous tumor fibers

“…#7500, CA, USA). Similar to previously reported methods 36 , GSC23 cells were transfected with red fluorescence protein (RFP) gene, and hMSCs were transfected with green fluorescence protein (GFP) gene. In another method, GSC23 cells were also transfected with a lentiviral vector GV348 (sequence element: Ubi-MCS-SV40-puromycin) containing LOXP-STOP-LOXP-RFP gene, and MSCs were transfected with a lentiviral vector GV348 containing a CRE enzyme gene.…”

Mesenchymal stem cells promote glioma neovascularization in vivo by fusing with cancer stem cells

“…Informed consent was obtained from the patient prior to sample acquisition.SU3 cells were regularly authenticated on the basis of morphology, expression of CD133 and nestin, sphere formation ability and tumorigenicity and were regularly examined for (absence of) mycoplasma by Mycoplasma Stain Assay Kit (Beotime, China). SU3 cells were transfected with the RFP gene using a lentiviral-mediated gene transfection kit (GeneChem, Shanghai, China) [6], and cultured in stem cell medium. Briefly, 20 ng/ml bFGF, 20 ng/ml EGF and 1 × B27 supplement were added to DMEM/F12.…”