title: First trimester embryo-fetoscopic and ultrasound-guided fetal blood sampling for ex vivo viral transduction of cultured human fetal mesenchymal stem cells.
abstract: BACKGROUND
  Intrauterine stem cell transplantation is a promising approach for early onset genetic diseases. However, its utility is limited by the development of the fetal immune system after 14 weeks gestation. An ex vivo gene therapy approach targeting autologous first trimester stem cells to replace the missing or defective gene product should overcome this barrier. We investigated the feasibility of harvesting circulating first trimester human fetal mesenchymal stem cells (hfMSCs) for ex vivo gene therapy.
  
  
  METHODS
  Thin-gauge embryofetoscopic-directed or ultrasound-guided blood sampling (FBS) was performed in 18 pre-termination fetuses at a mean of 10(+0) (range 7(+2) to 13(+4)) weeks gestation through extra-fetal vessels. Harvested blood was plated for isolation of hfMSC and transduced by lentiviruses.
  
  
  RESULTS
  FBS was successful in 12/18 procedures (67%). Success rates were comparable in fetoscopic (4/6) and ultrasound-guided (8/12) procedures, but procedural time was shorter in the ultrasound-guided arm (P = 0.01). Fetal bradycardia occurred post-FBS in 33% and 25% of fetoscopic and ultrasound cases, respectively, 5 min post-procedure. hfMSCs were isolated in two-thirds of cases, with high efficiency lentiviral transduction achieved without affecting short-term cell renewal.
  
  
  CONCLUSIONS
  This phase-one study demonstrates the feasibility of the ex vivo fetal gene therapy approach, in which harvested hfMSCs are genetically manipulated prior to infusion back into the fetus where they should engraft and home to injured tissues. The fetal ex vivo gene therapy paradigm is also of relevance to haemopoietic stem cells to treat inherited haematological diseases. Optimization of stem cell harvest and longer-term safety is required before translation into clinical trials in ongoing pregnancies.
authors.affiliation: Institute of Reproductive and Developmental Biology, Imperial College London, Hammersmith Campus, London, UK. jerrychan@nus.edu.sg
authors.forename: Jerry
authors.initials: J
authors.lastname: Chan
authors.forename: Sailesh
authors.initials: S
authors.lastname: Kumar
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Fisk
mesh_terms: D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D053686:Fetal Stem Cells / Q000166:cytology*; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D013234:Stem Cells; D014463:Ultrasonography / Q000379:methods