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Cell Strains Derived from Long‐term 3D Tumor Models of MCF‐7 Display Characteristics and Phenotypes Consistent with Stem‐cell Populations
Author(s) -
Topel Samantha,
Lyden Timothy
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.496.18
Subject(s) - trypsinization , flow cytometry , phenotype , cell , biology , stem cell , cell culture , mcf 7 , spheroid , microbiology and biotechnology , cancer cell , pathology , cancer , genetics , human breast , medicine , biochemistry , trypsin , gene , enzyme
In a continuing series of studies, our laboratory has been examining the cell populations associated with long‐term 3‐D artificial tumor models generated using standard MCF‐7 breast ductal adenocarcinoma cells. In these studies we have developed long‐term cultures and grown them out to 1, 2, 3 and 4 years respectively. This report focuses on MCF‐7 tumors developed on de‐cellularized and de‐calcified bovine trabecular bone scaffolds. These long‐term cultures generate populations of cells throughout the entire culture period by shedding both individual cells and cell clusters, often in the form of spheroids. These various shed cell populations then form colonies and eventually monolayers of cells, from which the scaffold must be removed to new wells as a form of “passage”. This entire process of monolayer formation then repeats itself again within a week in the new wells. In addition to shed cells we have also harvested cell strains directly from the scaffolds by standard trypsinization for 15 minutes followed by a second trypsinization of an addition 15 minutes. The paired cultures of rapidly released and slowly released cells display significantly different phenotypes and present with differential marker patterns as well. In addition, we have also established cell strains from both explanting cells directly associated with the artificial tumors and colonies which establish themselves at a distance from the original tumor. These distinct populations also display unique phenotypes and marker patterns by flow cytometry. In these analyses it is clear that at least two major populations occur in both types of monolayer, but that the relative proportion is distinctly different, with a much increased stem‐like cell population being observed in released and floating cells from the colony strain but far less so from the explanting strain. Based on both size and CD44/CD24 immunophenotyping, we have established that this distinct cell population is mainly associated with distant colonies. It is our working hypothesis that these respective populations represent differential metastatic states in artificial tumor‐derived cell strains. Furthermore, we contend that this work characterizing cell populations derived from long‐term tumor models further confirms our overall conclusion that these artificial tumor tissues provide an effective model of tumor development including progression and metastasis in‐vitro. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .