Mouse sarcoma L1 cell line holoclones have a stemness signature

Abstract

Accumulating data suggest that cancers contain a fraction of cells called cancer stem cells (CSCs), that may be responsible for upkeep and relapses of disease. In experimental settings, CSCs are regarded as most effective at tumour initiation in in vivo assays. Since the first isolation of cancer stem cells from acute myeloid leukaemia in 1994, cancer stem cells have been identified in human solid tumours and they have also been found in the established cell lines, based on ability of CSCs to form in vitro colonies of a specific morphology, called holoclones. Our study examined the ability of a mouse sarcoma cell line, derived from a lung metastasis of a BALB/c mouse and established as a stably growing line (L1), to produce holoclones in vitro. We aimed to verify a stemness signature of the holoclone cells. The L1 cell line was found to form holoclone colonies in vitro, which were shown to contain a percentage of CSC-like cells. A fraction of the L1 cells was able to repopulate the original cell line, and presented an increased clonogenic and metastatic potential (18th passage). In addition, MTT assay and flow cytometry of the side population fraction revealed that these cells were more resistant to chemotherapeutic drugs than the original cell line, and over-expressed the anti-apoptotic genes, GRP78 and GADD153. We conclude that mouse L1 sarcoma cell line contains CSC-like cells.

Heterogeneous Cell Types in Single-cell-derived Clones of MCF7 and MDA-MB-231 Cells

“…Also, the lowest efficiency of holoclone formation was found in MCF7-RFP cells. As The major characteristic of CSCs is to neutralize the effect of anticancer drugs via efflux transporter proteins (21,26). In this study, the holoclones of MCF7 and MDA-MB-231 cells showed resistance to Dox.…”

“…Multidrug resistance assay. Single-cell-derived holoclones were treated with Dox (1.5 μg/ml) for 24 h as reported (21). Control and treated clones were fixed as described and nuclei were stained with 4' 6-diamidino-2-phenylindole dihydrochloride (DAPI; 5 μg/ml) for 20 min in dark.…”

Prenyl Ammonium Salts – New Carriers for Gene Delivery: A B16-F10 Mouse Melanoma Model

“…B16-F10 mouse melanoma (ATCC ® CRL-6475 ™ ) and L1 mouse sarcoma cells derived from a lung metastasis of a BALB/c mouse [ 20 ] were cultured in Dulbecco’s Modified Eagle Medium (Gibco BRL, Life Technologies) supplemented with 10% FBS (Lonza, BioWhittaker). Cells were maintained at 37°C in a humidified 5% CO 2 atmosphere with media changes every third day.…”

Molecular Chaperones in Cancer Stem Cells: Determinants of Stemness and Potential Targets for Antitumor Therapy

“…There were reports about the high intracellular GRP78 levels in CSC-like cells of different origins [206][207][208]212]. In 2010, Wu et al demonstrated that knockdown of GRP78 in tumor-initiating (CSC-like) cells from head and neck cancer impairs their self-renewing capacity, tumorigenic potential, and expression of stemness-related genes, while reducing the SP cell's proportion and enhancing apoptosis [213].…”