Isolation and Establishment of Human Tumor Stem Cells

Roberts, Penelope E.

doi:10.1016/s0091-679x(08)00015-0

Cited by 22 publications

(18 citation statements)

References 10 publications

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“…CCA stem-like cells were identified by using a functional tool of 3D culture system based on defined serum-free medium, as shown in many patient-derived tumors [21,22]. First, established (SG231, HUCCT1, CCLP1) [23–25] and primary human intrahepatic CCA-derived (CCA4) cell lines [26] were tested for sphere-forming efficiency (SFE) as a representation of tumor stem-like subset.…”

Section: Resultsmentioning

confidence: 99%

Cholangiocarcinoma stem-like subset shapes tumor-initiating niche by educating associated macrophages

Raggi

Correnti

Sica

et al. 2017

Journal of Hepatology

144

109

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Background & Aims A therapeutically challenging subset of cells, termed cancer stem cells (CSCs) are responsible for cholangiocarcinoma (CCA) clinical severity. Presence of tumor-associated macrophages (TAMs) has prognostic significance in CCA and other malignancies. Thus, we hypothesized that CSCs may actively shape their tumor-supportive immune niche. Methods CCA cells were cultured in 3D conditions to generate spheres. CCA sphere analysis of in vivo tumorigenic-engraftment in immune-deficient mice and molecular characterization was performed. The in vitro and in vivo effect of CCA spheres on macrophage precursors was tested after culturing healthy donor cluster of differentiation (CD)14+ with CCA-sphere conditioned medium. Results CCA spheres engrafted in 100% of transplanted mice and revealed a significant 20.3-fold increase in tumor-initiating fraction (p = 0.0011) and a sustained tumorigenic potential through diverse xenograft-generations. Moreover, CCA spheres were highly enriched for CSC, liver cancer and embryonic stem cell markers both at gene and protein levels. Next, fluorescence-activated cell sorting analysis showed that in the presence of CCA sphere conditioned medium, CD14+ macrophages expressed key markers (CD68, CD115, human leukocyte antigen-D related, CD206) indicating that CCA sphere conditioned medium was a strong macrophage-activator. Gene expression profile of CCA sphere activated macrophages revealed unique molecular TAM-like features confirmed by high invasion capacity. Also, freshly isolated macrophages from CCA resections recapitulated a similar molecular phenotype of in vitro-educated macrophages. Consistent with invasive features, the largest CD163+ set was found in the tumor front of human CCA specimens (n = 23) and correlated with a high level of serum cancer antigen 19.9 (n = 17). Among mediators released by CCA spheres, only interleukin (IL)13, IL34 and osteoactivin were detected and further confirmed in CCA patient sera (n = 12). Surprisingly, a significant association of IL13, IL34 and osteoactivin with sphere stem-like genes was provided by a CCA database (n = 104). In vitro combination of IL13, IL34, osteoactivin was responsible for macrophage-differentiation and invasion, as well as for in vivo tumor-promoting effect. Conclusion CCA-CSCs molded a specific subset of stem-like associated macrophages thus providing a rationale for a synergistic therapeutic strategy for CCA-disease. Lay summary Immune plasticity represents an important hallmark of tumor outcome. Since cancer stem cells are able to manipulate stromal cells to their needs, a better definition of the key dysregulated immune subtypes responsible for cooperating in supporting tumor initiation may facilitate the development of new therapeutic approaches. Considering that human cholangiocarcinoma represents a clinical emergency, it is essential to move to predictive models in order to understand the adaptive process of macrophage component (imprinting, polarization and maintenance) engaged by tumor stem-like ...

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Section: Resultsmentioning

confidence: 99%

Cholangiocarcinoma stem-like subset shapes tumor-initiating niche by educating associated macrophages

Raggi

Correnti

Sica

et al. 2017

Journal of Hepatology

144

109

View full text Add to dashboard Cite

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“…To identify cell-surface cancer antigens suitable for mAbbased targeting, we have characterized mAbs generated in a target-unbiased fashion from intact cell immunizations of serum-free adapted tissue progenitor cells or cancer cell lines, including those exhibiting cancer stem cell properties (8,9). Immunohistochemical profiling of these mAbs reveals those displaying differential expression on human cancer tissues compared with normal tissue, whereas biochemical analyses identify the cell-surface cancer antigen recognized (10).…”

Section: Introductionmentioning

confidence: 99%

Development of an Fc-Enhanced Anti–B7-H3 Monoclonal Antibody with Potent Antitumor Activity

Loo

Alderson

Chen

et al. 2012

Clinical Cancer Research

230

171

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Purpose: The goal of this research was to harness a monoclonal antibody (mAb) discovery platform to identify cell-surface antigens highly expressed on cancer and develop, through Fc optimization, potent mAb therapies toward these tumor-specific antigens.Experimental Design: Fifty independent mAbs targeting the cell-surface immunoregulatory B7-H3 protein were obtained through independent intact cell-based immunizations using human tissue progenitor cells, cancer cell lines, or cell lines displaying cancer stem cell properties. Binding studies revealed this natively reactive B7-H3 mAb panel to bind a range of independent B7-H3 epitopes. Immunohistochemical analyses showed that a subset displayed strong reactivity to a broad range of human cancers while exhibiting limited binding to normal human tissues. A B7-H3 mAb displaying exquisite tumor/normal differential binding was selected for humanization and incorporation of an Fc domain modified to enhance effectormediated antitumor function via increased affinity for the activating receptor CD16A and decreased binding to the inhibitory receptor CD32B.Results: MGA271, the resulting engineered anti-B7-H3 mAb, mediates potent antibody-dependent cellular cytotoxicity against a broad range of tumor cell types. Furthermore, in human CD16A-bearing transgenic mice, MGA271 exhibited potent antitumor activity in B7-H3-expressing xenograft models of renal cell and bladder carcinoma. Toxicology studies carried out in cynomolgus monkeys revealed no significant test article-related safety findings.Conclusions: This data supports evaluation of MGA271 clinical utility in B7-H3-expressing cancer, while validating a combination of a nontarget biased approach of intact cell immunizations and immunohistochemistry to identify novel cancer antigens with Fc-based mAb engineering to enable potent antitumor activity.

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“…The strength of this method is the ability to define the correct environment (niche) required to maintain the stemness of the CSCs. This method has been successfully used to select and expand cell populations with CSC characteristics from patients with colon and lung cancers [12] and gliomas [13]. In this study, we developed a defined serum-free medium that is able to enrich and expand HCSCs, maintain their stem cell traits but are still capable of tumor initiation.…”

Section: Introductionmentioning

confidence: 99%

Establishment of a novel system for the culture and expansion of hepatic stem-like cancer cells

Jiang

Wang

et al. 2015

Cancer Letters

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Hepatocellular carcinoma (HCC) is a major primary liver malignancy in adults. Despite the progress made, the outcome of the treatment to this disease is less than satisfactory as the post therapy tumor recurrence is almost inevitable. Accumulating pieces of evidence have suggested that the recurrence is due to the existence of a subpopulation of the HCC cells that possess the properties of stem cells and are resistant to radiation and chemotherapy. It is therefore important to understand the characteristics of this subpopulation of HCC cells, and which requires the establishment of an in vitro system to study these stem-like cancer cells. However, despite extensive efforts, the progress in establishing such an in vitro system has been slow largely due to the lack of definitive biomarkers in the isolation and expansion of these cells. In order to successfully maintain and expand HCC CSCs, we first optimized the culture system. We establish a novel medium system that allows the culture and enrichment of these hepatic stem-like cancer cells from both hepatoma cells and human primary HCC cells. These cells exhibited typical stem cell properties, such as enhanced stem cell markers, gain of EMT properties and drug resistance, and more importantly, stronger tumor-initiating capabilities. The medium may help to establish an in vitro model for hepatic cancer stem cell (HCSC) studies, which may contribute to the development of novel cell therapies and new drugs for the treatment of HCC.

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Isolation and Establishment of Human Tumor Stem Cells

Cited by 22 publications

References 10 publications

Cholangiocarcinoma stem-like subset shapes tumor-initiating niche by educating associated macrophages

Cholangiocarcinoma stem-like subset shapes tumor-initiating niche by educating associated macrophages

Development of an Fc-Enhanced Anti–B7-H3 Monoclonal Antibody with Potent Antitumor Activity

Establishment of a novel system for the culture and expansion of hepatic stem-like cancer cells

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