Autologous Transplantation of Amniotic Fluid-Derived Mesenchymal Stem Cells into Sheep Fetuses

Cell Transplantation

Article

H. Bobby Gaspar¹  Waseem Qasim¹  Anna L. David²  Khalil Abi Nader²  Vedanta Mehta²  Paolo De Coppi²  Elisa Filppi²  S. W. Steven Shaw³  Annalisa Gastadello⁴  Sveva Bollini⁴  Mara Cananzi⁴ 

[1] Department of Paediatric Immunology, Institute of Child Health, University College London, London, UK;Prenatal Cell and Gene Therapy Group, Institute for Women's Health, University College London, London, UK;Prenatal Cell and Gene Therapy Group, Institute for Women's Health, University College London, London, UK; Surgery Unit, Institute of Child Health, University College London, London, UK; Department of Obstetrics and Gynaecology, Chang Gung Memorial Hospital at Linkou and Chang Gung University, College of Medicine, Taoyuan, Taiwan;Surgery Unit, Institute of Child Health, University College London, London, UK;

关键词: Autologous stem cell transplantation;    Amniotic fluid stem cell;    Sheep;    In utero;   

DOI  :  10.3727/096368910X543402

 received in 2010-02-22, accepted in 2010-11-08,  发布年份 2011

来源: Sage Journals

【 摘 要 】

Long-term engraftment and phenotype correction has been difficult to achieve in humans after in utero stem cell transplantation mainly because of allogeneic rejection. Autologous cells could be obtained during gestation from the amniotic fluid with minimal risk for the fetus and the mother. Using a sheep model, we explored the possibility of using amniotic fluid mesenchymal stem cells (AFMSCs) for autologous in utero stem cell/gene therapy. We collected amniotic fluid (AF) under ultrasound-guided amniocentesis in early gestation pregnant sheep (n = 9, 58 days of gestation, term = 145 days). AFMSCs were isolated and expanded in all sampled fetal sheep. Those cells were transduced using an HIV vector encoding enhanced green fluorescent protein (GFP) with 63.2% (range 38.3–96.2%) transduction efficiency rate. After expansion, transduced AFMSCs were injected into the peritoneal cavity of each donor fetal sheep at 76 days under ultrasound guidance. One ewe miscarried twin fetuses after amniocentesis. Intraperitoneal injection was successful in the remaining 7 fetal sheep giving a 78% survival for the full procedure. Tissues were sampled at postmortem examination 2 weeks later. PCR analysis detected GFP-positive cells in fetal tissues including liver, heart, placenta, membrane, umbilical cord, adrenal gland, and muscle. GFP protein was detected in these tissues by Western blotting and further confirmed by cytofluorimetric and immunofluorescence analyses. This is the first demonstration of autologous stem cell transplantation in the fetus using AFMSCs. Autologous cells derived from AF showed widespread organ migration and could offer an alternative way to ameliorate prenatal congenital disease.

【 授权许可】

Unknown   
© 2011 Cognizant Comm. Corp.

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