Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion

Donatella Amendola, Marta Nardella, Loredana Guglielmi, Lidia Cerquetti, Elisabetta Carico, Viola Alesi, Manuela Porru, Carlo Leonetti, Claudia Bearzi, Roberto Rizzi, Igea D'Agnano, Antonio Stigliano, Giuseppe Novelli, Barbara Bucci

Research output: Contribution to journalArticle

Abstract

The cancer stem cell model links neoplastic cells with normal stem cell biology, but little is known on how normal stem cells are transformed into cancer stem cells. Methods: To investigate the processes underlying the transformation of normal stem cells we developed in vitro a cancer stem cell model from human amniotic and chorionic placenta membranes. In this model we studied the expression of specific stem cell molecules by flow cytometry, and genes, by real time RT-PCR. Microscopy immunfluorescence was employed to investigate the proliferative and differentiation patterns. Fluorescence microscopy and FACS were employed to investigate the proliferative and differentiation patterns. To evaluate the tumorigenic potential of our model we injected the cells into NOD.CB17-Prkdcscid/NCrHsd mice. Results: Normal human stem cells from amniotic and chorionic placenta membranes were converted into neural cell lineages, under specific conditions, to form secondary neurospheres with a capacity for self-renewal. After extensive in vitro culture, these cells underwent spontaneous transformations and acquired a neuroblastoma (NB)-like phenotype with an elevated proliferative potential that is comparable to established neuroblastoma cell lines. The ability of these cells to transform their phenotype was evidenced by increased clonogenic ability in vitro; by augmented expression level of certain proliferation- and transformation-related genes (e.g., CCNA2, MYCN, ENPP2, GRIA3, and KIT); by the presence of multinucleated and hyperdiploid cells. We further demonstrated that the transformed phenotype is an NB by measuring the expression of NB-specific markers, disialoganglioside GD2 and N-Myc proteins. Conclusions: We have developed a cancer stem cell model starting from normal human stem cells derived from amniotic and chorionic placenta membranes. These cells are able to differentiate into neural cell lineages and to undergo spontaneous transformations and acquire an NB-like phenotype.

Original languageEnglish
Article number55
JournalStem Cell Research and Therapy
Volume5
Issue number2
DOIs
Publication statusPublished - Apr 22 2014

Fingerprint

Stem cells
Placenta
Neuroblastoma
Stem Cells
Neoplastic Stem Cells
Phenotype
Cell Lineage
Membranes
Proto-Oncogene Proteins c-myc
Polyploidy
Fluorescence Microscopy
Genes
Cell Biology
In Vitro Techniques
Real-Time Polymerase Chain Reaction
Cytology
Microscopy
Flow Cytometry
Cell Culture Techniques
Flow cytometry

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Molecular Medicine
  • Cell Biology
  • Medicine (miscellaneous)
  • Medicine(all)

Cite this

Amendola, D., Nardella, M., Guglielmi, L., Cerquetti, L., Carico, E., Alesi, V., ... Bucci, B. (2014). Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion. Stem Cell Research and Therapy, 5(2), [55]. https://doi.org/10.1186/scrt444

Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion. / Amendola, Donatella; Nardella, Marta; Guglielmi, Loredana; Cerquetti, Lidia; Carico, Elisabetta; Alesi, Viola; Porru, Manuela; Leonetti, Carlo; Bearzi, Claudia; Rizzi, Roberto; D'Agnano, Igea; Stigliano, Antonio; Novelli, Giuseppe; Bucci, Barbara.

In: Stem Cell Research and Therapy, Vol. 5, No. 2, 55, 22.04.2014.

Research output: Contribution to journalArticle

Amendola, D, Nardella, M, Guglielmi, L, Cerquetti, L, Carico, E, Alesi, V, Porru, M, Leonetti, C, Bearzi, C, Rizzi, R, D'Agnano, I, Stigliano, A, Novelli, G & Bucci, B 2014, 'Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion', Stem Cell Research and Therapy, vol. 5, no. 2, 55. https://doi.org/10.1186/scrt444
Amendola, Donatella ; Nardella, Marta ; Guglielmi, Loredana ; Cerquetti, Lidia ; Carico, Elisabetta ; Alesi, Viola ; Porru, Manuela ; Leonetti, Carlo ; Bearzi, Claudia ; Rizzi, Roberto ; D'Agnano, Igea ; Stigliano, Antonio ; Novelli, Giuseppe ; Bucci, Barbara. / Human placenta-derived neurospheres are susceptible to transformation after extensive in vitro expansion. In: Stem Cell Research and Therapy. 2014 ; Vol. 5, No. 2.
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abstract = "The cancer stem cell model links neoplastic cells with normal stem cell biology, but little is known on how normal stem cells are transformed into cancer stem cells. Methods: To investigate the processes underlying the transformation of normal stem cells we developed in vitro a cancer stem cell model from human amniotic and chorionic placenta membranes. In this model we studied the expression of specific stem cell molecules by flow cytometry, and genes, by real time RT-PCR. Microscopy immunfluorescence was employed to investigate the proliferative and differentiation patterns. Fluorescence microscopy and FACS were employed to investigate the proliferative and differentiation patterns. To evaluate the tumorigenic potential of our model we injected the cells into NOD.CB17-Prkdcscid/NCrHsd mice. Results: Normal human stem cells from amniotic and chorionic placenta membranes were converted into neural cell lineages, under specific conditions, to form secondary neurospheres with a capacity for self-renewal. After extensive in vitro culture, these cells underwent spontaneous transformations and acquired a neuroblastoma (NB)-like phenotype with an elevated proliferative potential that is comparable to established neuroblastoma cell lines. The ability of these cells to transform their phenotype was evidenced by increased clonogenic ability in vitro; by augmented expression level of certain proliferation- and transformation-related genes (e.g., CCNA2, MYCN, ENPP2, GRIA3, and KIT); by the presence of multinucleated and hyperdiploid cells. We further demonstrated that the transformed phenotype is an NB by measuring the expression of NB-specific markers, disialoganglioside GD2 and N-Myc proteins. Conclusions: We have developed a cancer stem cell model starting from normal human stem cells derived from amniotic and chorionic placenta membranes. These cells are able to differentiate into neural cell lineages and to undergo spontaneous transformations and acquire an NB-like phenotype.",
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AU - Amendola, Donatella

AU - Nardella, Marta

AU - Guglielmi, Loredana

AU - Cerquetti, Lidia

AU - Carico, Elisabetta

AU - Alesi, Viola

AU - Porru, Manuela

AU - Leonetti, Carlo

AU - Bearzi, Claudia

AU - Rizzi, Roberto

AU - D'Agnano, Igea

AU - Stigliano, Antonio

AU - Novelli, Giuseppe

AU - Bucci, Barbara

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N2 - The cancer stem cell model links neoplastic cells with normal stem cell biology, but little is known on how normal stem cells are transformed into cancer stem cells. Methods: To investigate the processes underlying the transformation of normal stem cells we developed in vitro a cancer stem cell model from human amniotic and chorionic placenta membranes. In this model we studied the expression of specific stem cell molecules by flow cytometry, and genes, by real time RT-PCR. Microscopy immunfluorescence was employed to investigate the proliferative and differentiation patterns. Fluorescence microscopy and FACS were employed to investigate the proliferative and differentiation patterns. To evaluate the tumorigenic potential of our model we injected the cells into NOD.CB17-Prkdcscid/NCrHsd mice. Results: Normal human stem cells from amniotic and chorionic placenta membranes were converted into neural cell lineages, under specific conditions, to form secondary neurospheres with a capacity for self-renewal. After extensive in vitro culture, these cells underwent spontaneous transformations and acquired a neuroblastoma (NB)-like phenotype with an elevated proliferative potential that is comparable to established neuroblastoma cell lines. The ability of these cells to transform their phenotype was evidenced by increased clonogenic ability in vitro; by augmented expression level of certain proliferation- and transformation-related genes (e.g., CCNA2, MYCN, ENPP2, GRIA3, and KIT); by the presence of multinucleated and hyperdiploid cells. We further demonstrated that the transformed phenotype is an NB by measuring the expression of NB-specific markers, disialoganglioside GD2 and N-Myc proteins. Conclusions: We have developed a cancer stem cell model starting from normal human stem cells derived from amniotic and chorionic placenta membranes. These cells are able to differentiate into neural cell lineages and to undergo spontaneous transformations and acquire an NB-like phenotype.

AB - The cancer stem cell model links neoplastic cells with normal stem cell biology, but little is known on how normal stem cells are transformed into cancer stem cells. Methods: To investigate the processes underlying the transformation of normal stem cells we developed in vitro a cancer stem cell model from human amniotic and chorionic placenta membranes. In this model we studied the expression of specific stem cell molecules by flow cytometry, and genes, by real time RT-PCR. Microscopy immunfluorescence was employed to investigate the proliferative and differentiation patterns. Fluorescence microscopy and FACS were employed to investigate the proliferative and differentiation patterns. To evaluate the tumorigenic potential of our model we injected the cells into NOD.CB17-Prkdcscid/NCrHsd mice. Results: Normal human stem cells from amniotic and chorionic placenta membranes were converted into neural cell lineages, under specific conditions, to form secondary neurospheres with a capacity for self-renewal. After extensive in vitro culture, these cells underwent spontaneous transformations and acquired a neuroblastoma (NB)-like phenotype with an elevated proliferative potential that is comparable to established neuroblastoma cell lines. The ability of these cells to transform their phenotype was evidenced by increased clonogenic ability in vitro; by augmented expression level of certain proliferation- and transformation-related genes (e.g., CCNA2, MYCN, ENPP2, GRIA3, and KIT); by the presence of multinucleated and hyperdiploid cells. We further demonstrated that the transformed phenotype is an NB by measuring the expression of NB-specific markers, disialoganglioside GD2 and N-Myc proteins. Conclusions: We have developed a cancer stem cell model starting from normal human stem cells derived from amniotic and chorionic placenta membranes. These cells are able to differentiate into neural cell lineages and to undergo spontaneous transformations and acquire an NB-like phenotype.

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