Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Smith, Barry H.; Gazda, Lawrence S.; Conn, Bryan L.; Jain, Kanti; Asina, Shirin; Levine, Daniel M.; Parker, Thomas S.; Laramore, Melissa A.; Martis, Prithy C.; Vinerean, Horatiu V.; David, Eric M.; Qiu, Suizhen; Cordon‐Cardo, Carlos; Hall, Richard D.; Gordon, Bruce R.; Diehl, Carolyn H.; Stenzel, Kurt H.; Rubin, Albert L.

doi:10.1158/0008-5472.can-10-2254

Cited by 48 publications

(50 citation statements)

References 40 publications

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“…8 In another study, encapsulation of neoplastic cells in a growth-restricting hydrogel composed of agarose matrix leads to the selection of a stem cell subpopulation and the formation of tumorspheres. 9 However, our work demonstrates for the first time that CSCs can be maintained in the inert microenvironment of synthetic PEG hydrogels and the elastic modulus of the gel affects the growth of tumorspheres. The growth and maintenance of CSCs in other synthetic gels, such as polyhydroxyethyl methacrylate, polyvinylpolypyrrolidone, and polyvinyl alcohol, is of interest and will be investigated in future works.…”

Section: Discussionmentioning

confidence: 77%

“…6,7 Alginate and agarose gels are also used as a matrix to study the behavior of breast cancer cells under a 3D condition. 8,9 However, it is difficult to isolate and study cell response to individual factors in the microenvironment with naturally derived matrices that have many interactions with cell surface receptors. 10 In that regard, synthetic gels provide enormous flexibility in designing 3D matrices with a wide range of mechanical, physical, and biological properties that are crucial to the maintenance, differentiation, maturation, and fate of the encapsulated cells.…”

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confidence: 99%

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Three-Dimensional-Engineered Matrix to Study Cancer Stem Cells and Tumorsphere Formation: Effect of Matrix Modulus

Yang

Sarvestani²,

Moeinzadeh³

et al. 2013

Tissue Engineering Part A

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Maintenance of cancer stem cells (CSCs) is regulated by the tumor microenvironment. Synthetic hydrogels provide the flexibility to design three-dimensional (3D) matrices to isolate and study individual factors in the tumor microenvironment. The objective of this work was to investigate the effect of matrix modulus on tumorsphere formation by breast cancer cells and maintenance of CSCs in an inert microenvironment without the interference of other factors. In that regard, 4T1 mouse breast cancer cells were encapsulated in inert polyethylene glycol diacrylate hydrogels and the effect of matrix modulus on tumorsphere formation and expression of CSC markers was investigated. The gel modulus had a strong effect on tumorsphere formation and the effect was bimodal. Tumorsphere formation and expression of CSC markers peaked after 8 days of culture. At day 8, as the matrix modulus was increased from 2.5 kPa to 5.3, 26.1, and 47.1 kPa, the average tumorsphere size changed from 37 -6 mm to 57 -6, 20 -4, and 12 -2 mm, respectively; cell number density in the gel changed from 0.8 -0.1 · 10 5 cells/mL to 1.7 -0.2 · 10 5 , 0.4 -0.1 · 10 5 , and 0.2 -0.1 · 10 5 cells/mL after initial encapsulation of 0.14 · 10 5 cells/mL; and the expression of CD44 breast CSC marker changed from 17 -4-fold to 38 -9-, 3 -1-, and 2 -1-fold increase compared with the initial level. Similar results were obtained with MCF7 human breast carcinoma cells. Mouse 4T1 and human MCF7 cells encapsulated in the gel with 5.3 kPa modulus formed the largest tumorspheres and highest density of tumorspheres, and had highest expression of breast CSC markers CD44 and ABCG2. The inert polyethylene glycol hydrogel can be used as a model-engineered 3D matrix to study the role of individual factors in the tumor microenvironment on tumorigenesis and maintenance of CSCs without the interference of other factors.

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Section: Discussionmentioning

confidence: 77%

mentioning

confidence: 99%

Three-Dimensional-Engineered Matrix to Study Cancer Stem Cells and Tumorsphere Formation: Effect of Matrix Modulus

Yang

Sarvestani²,

Moeinzadeh³

et al. 2013

Tissue Engineering Part A

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“…Also other types of tumors have been examined for CSCs (13)(14)(15)(16). Many studies had chosen specific cell surface markers such as CD19, CD20, CD24, CD44 and CD133 to characterize the subpopulation of CSCs.…”

Section: Introductionmentioning

confidence: 99%

Inhibition of CD147 expression by RNA interference reduces proliferation, invasion and increases chemosensitivity in cancer stem cell-like HT-29 cells

Chen

Pan

et al. 2015

International Journal of Oncology

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Abstract.The association between CD147 and cancer stem cells (CSCs) provides a new angle for cancer treatments. The aim of this study was to investigate the biological roles of CD147 in colorectal CSCs. The Oct4-green fluorescent protein (GFP) vector was used to isolate CSCs and pYr-mir30-shRNA was used to generate short hairpin RNA (shRNA) specifically for CD147. After RNA interference (RNAi), CD147 was evaluated by reverse transcription-quantitative PCR and western blot analysis, and its biological functions were assessed by MTT and invasion assays. The results showed that the differentiation of isolated CSC-like HT-29 cells was blocked and these cells were highly positive for CD44 and CD147. RNAi-mediated CD147 silencing reduced the expression of CD147 at both mRNA and protein levels. Moreover, the activities of proliferation and invasion were decreased obviously in CSCs. Knockdown of CD147 increased the chemosensitivity of CSC-like cells to gemcitabine, cisplatin, docetaxel at 0.1, 1 and 10 µM respectively, however, there was no significant difference among the three groups to paclitaxel at 10 µM. In conclusion, these results suggest that CD147 plays an important role in colorectal CSCs and might be regarded as a novel CSC-specific targeted strategy against colorectal cancer. IntroductionColorectal cancer, one of the three most common malignancies in the world, remains a major public health problem (1). In 2014, an estimated 71,830 men and 65,000 women were diagnosed with colorectal cancer and 26,270 men and 24,040 women will die of the disease (2). In recent years, remarkable advances have been made in colorectal cancer therapeutic approaches by using chemotherapy, radiotherapy, monoclonal antibodies and small-molecule inhibitors. However, the clinical outcome was far from expected as the effects of these improvements never continued for a long duration (3,4). Therefore, new and better therapies will be the key to reducing the incidence of colorectal cancer.A new emerging concept implicates that treatment failure occurs due to the existence of cancer stem cells (CSCs) that evade the treatment regimen (5,6). These cells had been characterized by pluripotency, self-renewal as well as tumorigenicity (7-9). CSCs were resistant to traditional therapies, leading to tumor recurrence and unpleasant prognosis for cancer patients (10,11). Conventional treatments mainly targeted the tumor cells, but not the CSCs. Thus, therapeutic strategies specifically targeting CSCs could eradicate colorectal cancer more effectively.CSCs were firstly identified in hematopoietic tumors by using cell surface and intracellular molecules (12). Also other types of tumors have been examined for CSCs (13-16). Many studies had chosen specific cell surface markers such as CD19, CD20, CD24, CD44 and CD133 to characterize the subpopulation of CSCs. Transcription factor Oct4 was also typically an intracellular marker (7,(17)(18)(19). Oct4 (also known as OCT3, OCT3/4) belongs to the POU (Pit-Oct-Unc) family of transcription factors. It can form...

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“…Using encapsulated tumor cells from one species to treat cancer in another species is attractive because strong immunologic (xenogeneic) barriers prevent cross-species seeding of unencapsulated cells. Beyond their potential therapeutic implications, the cancer macrobeads offer a unique opportunity to study the molecular biological basis of tumor cell proliferation control, in part, by virtue of their ability to select a subpopulation of cells that appear to have stem cell properties and to be responsible for enabling the founding and perpetuation of tumors (1). In addition, they provide a ready means for the in vitro preclinical testing of proposed therapeutic agents that, to be optimally effective, must show activity against such cells.…”

Section: Discussionmentioning

confidence: 99%

“…We have recently reported the ability of various tumor cell lines to form tumor colonies following encapsulation in agarose-agarose macrobeads (1). Tumor colonies arise in the core of the macrobeads from a rare population of the 150,000 cells originally encapsulated.…”

Section: Introductionmentioning

confidence: 99%

Hydrophilic Agarose Macrobead Cultures Select for Outgrowth of Carcinoma Cell Populations That Can Restrict Tumor Growth

et al. 2011

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Cancer cells and their associated tumors have long been considered to exhibit unregulated proliferation or growth. However, a substantial body of evidence indicates that tumor growth is subject to both positive and negative regulatory controls. Here, we describe a novel property of tumor growth regulation that is neither species nor tumor-type specific. This property, functionally a type of feedback control, is triggered by the encapsulation of neoplastic cells in a growth-restricting hydrogel composed of an agarose matrix with a second coating of agarose to form 6-to 8-mm diameter macrobeads. In a mouse cell model of renal adenocarcinoma (RENCA cells), this process resulted in selection for a stem cell-like subpopulation which together with at least one other cell subpopulation drove colony formation in the macrobeads. Cells in these colonies produced diffusible substances that markedly inhibited in vitro and in vivo proliferation of epithelial-derived tumor cells outside the macrobeads. RENCA cells in monolayer culture that were exposed to RENCA macrobead-conditioned media exhibited cell-cycle accumulation in S phase due to activation of a G 2 /M checkpoint. At least 10 proteins with known tumor suppression functions were identified by analysis of RENCA macrobead-conditioned media, the properties of which offer opportunities to further dissect the molecular basis for tumor growth control. More generally, macrobead culture may permit the isolation of cancer stem cells and other cells of the stem cell niche, perhaps providing strategies to define more effective biologically based clinical approaches to treat neoplastic disease. Cancer Res; 71(3); 725-35. Ó2011 AACR.

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Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Cited by 48 publications

References 40 publications

Three-Dimensional-Engineered Matrix to Study Cancer Stem Cells and Tumorsphere Formation: Effect of Matrix Modulus

Three-Dimensional-Engineered Matrix to Study Cancer Stem Cells and Tumorsphere Formation: Effect of Matrix Modulus

Inhibition of CD147 expression by RNA interference reduces proliferation, invasion and increases chemosensitivity in cancer stem cell-like HT-29 cells

Hydrophilic Agarose Macrobead Cultures Select for Outgrowth of Carcinoma Cell Populations That Can Restrict Tumor Growth

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