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However, levels of long-term hematopoietic engraftment achieved in experimental IUT to date are subtherapeutic, likely due to host fetal HSCs outcompeting their bone marrow (BM)-derived donor equivalents for space in the hematopoietic compartment. In the present study, we demonstrate that amniotic fluid stem cells (AFSCs; c-Kit+\/Lin\u2212) have hematopoietic characteristics and, thanks to their fetal origin, favorable proliferation kinetics in vitro and in vivo, which are maintained when the cells are expanded. IUT of autologous\/congenic freshly isolated or cultured AFSCs resulted in stable multilineage hematopoietic engraftment, far higher to that achieved with BM-HSCs. Intravascular IUT of allogenic AFSCs was not successful as recently reported after intraperitoneal IUT. Herein, we demonstrated that this likely due to a failure of timely homing of donor cells to the host fetal thymus resulted in lack of tolerance induction and rejection. This study reveals that intravascular IUT leads to a remarkable hematopoietic engraftment of AFSCs in the setting of autologous\/congenic IUT, and confirms the requirement for induction of central tolerance for allogenic IUT to be successful. Autologous, gene-engineered, and in vitro expanded AFSCs could be used as a stem cell\/gene therapy platform for the in utero treatment of inherited disorders of hematopoiesis. Stem Cells \u00a02019;37:1176\u20131188<\/jats:p>\n               <jats:sec>\n                  <jats:title\/>\n               <\/jats:sec>","DOI":"10.1002\/stem.3039","type":"journal-article","created":{"date-parts":[[2019,5,22]],"date-time":"2019-05-22T16:13:44Z","timestamp":1558541624000},"page":"1176-1188","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["In Utero Transplantation of Expanded Autologous Amniotic Fluid Stem Cells Results in Long-Term Hematopoietic Engraftment"],"prefix":"10.1093","volume":"37","author":[{"given":"Stavros P.","family":"Loukogeorgakis","sequence":"first","affiliation":[{"name":"a Stem Cells and Regenerative Medicine, Great Ormond Street Institute of Child Health, University College London, London, United Kingdom"},{"name":"b Center for Fetal Research, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2750-8405","authenticated-orcid":false,"given":"Panicos","family":"Shangaris","sequence":"additional","affiliation":[{"name":"a Stem Cells and Regenerative Medicine, Great Ormond Street Institute of Child Health, University College London, London, United Kingdom"},{"name":"c Research Department of Maternal and Fetal Medicine, Institute for Women's Health, University College London, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrica","family":"Bertin","sequence":"additional","affiliation":[{"name":"d Stem Cell and Regenerative Medicine Laboratory, Fondazione Instituto di Ricerca Pediatrica Citt\u00e0 della Speranza, University of Padova, Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Franzin","sequence":"additional","affiliation":[{"name":"d Stem Cell and Regenerative Medicine Laboratory, Fondazione Instituto di Ricerca Pediatrica Citt\u00e0 della Speranza, University of Padova, Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Piccoli","sequence":"additional","affiliation":[{"name":"d Stem Cell and Regenerative Medicine Laboratory, Fondazione Instituto di Ricerca Pediatrica Citt\u00e0 della Speranza, University of Padova, Padova, Italy"},{"name":"e Department of Women's and Children's Health, University of Padova, Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michela","family":"Pozzobon","sequence":"additional","affiliation":[{"name":"d Stem Cell and Regenerative Medicine Laboratory, Fondazione Instituto di Ricerca Pediatrica Citt\u00e0 della Speranza, University of Padova, Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sindhu","family":"Subramaniam","sequence":"additional","affiliation":[{"name":"a Stem Cells and Regenerative Medicine, Great Ormond Street Institute of Child Health, University College London, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfonso","family":"Tedeschi","sequence":"additional","affiliation":[{"name":"a Stem Cells and Regenerative Medicine, Great Ormond Street Institute of Child Health, University College London, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aimee G.","family":"Kim","sequence":"additional","affiliation":[{"name":"b Center for Fetal Research, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiying","family":"Li","sequence":"additional","affiliation":[{"name":"b Center for Fetal Research, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Camila G.","family":"Fachin","sequence":"additional","affiliation":[{"name":"b Center for Fetal Research, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA"},{"name":"f Federal University of S\u00e3o Paulo, S\u00e3o Paulo, Brazil"},{"name":"g Federal University of Paran\u00e1, Curitiba, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andre I. B. 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