A new tumorsphere culture condition restores potentials of self-renewal and metastasis of primary neuroblastoma in a mouse neuroblastoma model.
Tumorsphere culture enriches and expands tumor cells, thus providing important resources for cancer studies. However, as compared with metastatic tissues, primary tumors in the nervous system rarely give rise to long-surviving tumorspheres, thereby seriously limiting studies on these cancers. This might be due to the limited self-renewal capability of tumor cells and/or to inappropriate culture conditions. The growth and maintenance of tumor cells may depend on microenvironments and/or cell origins (e.g., primary or metastatic; stem cell-like or progenitor-like). Here, we attempted to establish a tumorsphere culture condition for primary neuroblastoma (NB). Primary tumors in MYCN transgenic mice, a NB model, could be serially transplanted, suggesting that these tumors contain cells with a high self-renewal potential. However, primary tumors did not give rise to tumorspheres under a serum-free neurosphere culture condition. The newly established culture condition (named PrimNeuS) contained two critical ingredients: fetal bovine serum and β-mercaptoethanol were essential for tumorsphere formation as well as indefinite passages. The spheres could be passaged more than 20 times without exhaustion under this condition, exhibited a property of differentiation and formed tumors in vivo. Unexpectedly, PrimNeuS revealed that the MYCN transgenic mice had bone marrow metastasis. Furthermore, subcutaneous tumors derived from tumorspheres of primary tumors showed bone marrow metastasis. Taken together, PrimNeuS provides resources for the study of NB and can be used as a powerful tool for the detection of minimal residual disease and for in vitro evaluation prior to personalized therapy.
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Dongliang
D
Cao
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Satoshi
S
Kishida
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Peng
P
Huang
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Ping
P
Mu
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Shoma
S
Tsubota
Center for Advanced Medicine and Clinical Research, Nagoya University Hospital. Nagoya, Japan.
Masaaki
M
Mizuno
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Kenji
K
Kadomatsu
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D060833:Cellular Microenvironment / Q000502:physiology*; D008623:Mercaptoethanol; D051379:Mice; D008822:Mice, Transgenic; D000071447:N-Myc Proto-Oncogene Protein; D009362:Neoplasm Metastasis / Q000503:physiopathology*; D009447:Neuroblastoma / Q000503:physiopathology*; D011518:Proto-Oncogene Proteins / Q000235:genetics; D044967:Serum; D014407:Tumor Cells, Cultured / Q000502:physiology*
C571825:MYCN protein, mouse; D000071447:N-Myc Proto-Oncogene Protein; D011518:Proto-Oncogene Proteins; D008623:Mercaptoethanol