In this article we used immunohistochemistry and FACS analyses to show that cells expressing markers typical of human stem cells such as SSEA4, OCT-4, ALP, and CD117 are present within the cytotrophoblastic tissue of human fetal chorionic villus samples (CVSs). After immunoselection of CV cells for SSEA4, FACS analyses showed an increased number of cells positive for OCT-4 and ALP and a small percentage (around 4%) of side population (SP) cells. In the same cell population, RT-PCR indicated the presence of OCT-4, NANOG, and SOX2 transcripts, also typical of stem cells. Depending on the in vitro conditions, a subset of SSEA4+ cells formed colonies resembling hESCs, with limited self renewal ability. At the same time, these cells were able to differentiate in vitro into derivatives of all three germ layers. When inoculated into immunocompromised mice, SSEA4+ cells did not form teratomas but were able to populate depleted hematopoietic tissues. Moreover, after injection into mouse blastocysts, they were incorporated into the inner cell mass and could be traced into several tissues of the adult chimeric mice. Finally, we show that SSEA4+ cells isolated from fetuses affected by Spinal Muscular Atrophy (SMA) can be genetically corrected with high efficiency in culture by Small Fragment Homologous Recombination (SFHR), a gene targeting approach. Taken together, our results indicate that SSEA4+ cells obtained from human CVSs contain a subpopulation of multipotent cells that we propose to name Human Cytotrophoblastic-derived Multipotent Cells (hCTMCs). These cells may be a safe and convenient source of cells for cell-based therapy, as well as an ideal target for in utero fetal gene therapy.

Spitalieri, P., Cortese, G., Pietropolli, A., Filareto, A., Dolci, S., Klinger, F.g., et al. (2009). Identification of multipotent cytotrophoblast cells from human first trimester chorionic villi. CLONING AND STEM CELLS, 11(4), 535-556 [10.1089/clo.2009.0046].

Identification of multipotent cytotrophoblast cells from human first trimester chorionic villi

SPITALIERI, PAOLA;PIETROPOLLI, ADALGISA;Dolci, S;KLINGER, FRANCESCA GIOIA;GIARDINA, EMILIANO;DI CESARE, SILVIA;LAURO, DAVIDE;NOVELLI, GIUSEPPE;DE FELICI, MASSIMO;SANGIUOLO, FEDERICA CARLA
2009-12-01

Abstract

In this article we used immunohistochemistry and FACS analyses to show that cells expressing markers typical of human stem cells such as SSEA4, OCT-4, ALP, and CD117 are present within the cytotrophoblastic tissue of human fetal chorionic villus samples (CVSs). After immunoselection of CV cells for SSEA4, FACS analyses showed an increased number of cells positive for OCT-4 and ALP and a small percentage (around 4%) of side population (SP) cells. In the same cell population, RT-PCR indicated the presence of OCT-4, NANOG, and SOX2 transcripts, also typical of stem cells. Depending on the in vitro conditions, a subset of SSEA4+ cells formed colonies resembling hESCs, with limited self renewal ability. At the same time, these cells were able to differentiate in vitro into derivatives of all three germ layers. When inoculated into immunocompromised mice, SSEA4+ cells did not form teratomas but were able to populate depleted hematopoietic tissues. Moreover, after injection into mouse blastocysts, they were incorporated into the inner cell mass and could be traced into several tissues of the adult chimeric mice. Finally, we show that SSEA4+ cells isolated from fetuses affected by Spinal Muscular Atrophy (SMA) can be genetically corrected with high efficiency in culture by Small Fragment Homologous Recombination (SFHR), a gene targeting approach. Taken together, our results indicate that SSEA4+ cells obtained from human CVSs contain a subpopulation of multipotent cells that we propose to name Human Cytotrophoblastic-derived Multipotent Cells (hCTMCs). These cells may be a safe and convenient source of cells for cell-based therapy, as well as an ideal target for in utero fetal gene therapy.
dic-2009
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore MED/03 - GENETICA MEDICA
Settore MED/13 - ENDOCRINOLOGIA
Settore MED/50 - SCIENZE TECNICHE MEDICHE APPLICATE
Settore MED/09 - MEDICINA INTERNA
English
Con Impact Factor ISI
Fetus; Animals; Chorionic Villi; Proto-Oncogene Proteins c-kit; Humans; Germ Layers; Chimera; Muscular Atrophy, Spinal; Chemokine CCL27; Homeodomain Proteins; Cell Differentiation; Trophoblasts; Mice; Gene Therapy; SOXB1 Transcription Factors; Pregnancy; Octamer Transcription Factor-3; Multipotent Stem Cells; Pregnancy Trimester, First; Cells, Cultured; Hematopoietic System; Stage-Specific Embryonic Antigens; Gene Targeting; Teratoma; Female
Spitalieri, P., Cortese, G., Pietropolli, A., Filareto, A., Dolci, S., Klinger, F.g., et al. (2009). Identification of multipotent cytotrophoblast cells from human first trimester chorionic villi. CLONING AND STEM CELLS, 11(4), 535-556 [10.1089/clo.2009.0046].
Spitalieri, P; Cortese, G; Pietropolli, A; Filareto, A; Dolci, S; Klinger, Fg; Giardina, E; DI CESARE, S; Bernardini, L; Lauro, D; Scaldaferri, M; Sca...espandi
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/41961
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