Identification of multipotent cytotrophoblast cells from human first trimester chorionic villi.
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
Paola
P
Department of Biopathology, Genetics Unit, Tor Vergata University of Rome, via Montpellier, Italy.
Cortese
Giancarlo
G
Pietropolli
Adalgisa
A
Filareto
Antonio
A
Dolci
Susanna
S
Klinger
Francesca Gioia
FG
Giardina
Emiliano
E
Di Cesare
Silvia
S
Bernardini
Laura
L
Lauro
Davide
D
Scaldaferri
M Lucia
ML
Scaldaferri
H Lucia
HL
Citro
Gennaro
G
Novelli
Giuseppe
G
De Felici
Massimo
M
Sangiuolo
Federica
F
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D054425:Chemokine CCL27 / Q000378:metabolism*; D002678:Chimera / Q000235:genetics / Q000378:metabolism; D002824:Chorionic Villi; D005260:Female; D005333:Fetus / Q000166:cytology / Q000502:physiology*; D018390:Gene Targeting; D015316:Genetic Therapy; D005855:Germ Layers / Q000166:cytology; D006413:Hematopoietic System / Q000166:cytology; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D009134:Muscular Atrophy, Spinal / Q000235:genetics / Q000628:therapy; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D055748:SOXB1 Transcription Factors / Q000235:genetics; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism*; D013724:Teratoma / Q000473:pathology; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation
C492276:CCL27 protein, human; D054425:Chemokine CCL27; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D055746:Stage-Specific Embryonic Antigens; C066584:stage-specific embryonic antigen-4; D019009:Proto-Oncogene Proteins c-kit