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• W2003641451 abstract "A major obstacle for successful in-utero gene therapy with stem cells is defining the optimal site, timing and method for delivery of stem cells. Sonography has enabled in-vivo observation of the normal changes with time of the gestational sac compartments1-4 and sampling of amniotic and extra-embryonic celomic fluids in humans and non-human primates5, 6. Investigators from King's College in London were the first to report on the feasibility of ultrasound-guided celocentesis in humans prior to elective termination of pregnancy5. Since then, celocentesis has: provided insights into the biochemical composition of celomic fluid7, 8; enabled the prenatal diagnosis of genetic disorders in very early pregnancy9; and opened the doors to experimental in-utero stem-cell therapy10. If replacement of celomic fluid with specific culture medium does not cause a miscarriage, the celocentesis procedure could also be used to condition the celomic cavity prior to or during in-utero stem-cell therapy to enhance survival and the potential for engraftment of injected stem cells. We describe three cases of partial replacement of celomic fluid by hematopoietic stem-cell culture medium in a baboon model. Animal studies were approved by the Animal Care Committee at the University of Oklahoma. The culture medium injected consisted of RPMI-1640 supplemented with 10% fetal bovine serum (Sigma, St. Louis, MO, USA) and 25 ng/mL stem-cell factor (SCF, R&D Systems, Mineapolis, MN, USA), 25 ng/mL FLT-3 ligand (FLTeL, R&D Systems), 10 ng/mL thrombopoietin (TPO, Sigma) and 10 ng/mL IL-6 (IL6, Sigma). The percentage of extracelomic fluid (EECF) exchanged was calculated by subtracting the volume of the amniotic sac and yolk sac from that determined for the gestational sac (volumes for each cavity calculated with the formulae 4/3πr3). Fetuses were followed beyond 6–11 months to evaluate the outcome. In the first animal, 1.5 mL celomic fluid was replaced with 1.5 mL culture medium (i.e. the amount of EECF exchanged with culture medium was c. 10%) at 42 days post-conception (term pregnancy in baboons, 178–182 days). In the second animal, 3 mL celomic fluid was replaced with 3 mL culture medium (c. 30%) at 38 days post-conception, and in the third animal, 5 mL celomic fluid was replaced with 5 mL culture medium (c. 45%) at 41 days post-conception. All animals delivered healthy newborns at term and at 6–11 months of age the infants were developing normally according to institutionally derived nomograms. These findings are important because they provide a procedure that can enable aspiration of greater amounts of extracelomic fluid as previously reported for very early prenatal diagnosis9, and they suggest that replacement of celomic fluid with specific culture medium could be used to condition the extra-embryonic celomic cavity prior to or during in-utero stem-cell therapy in the pre-immune embryo. This conditioning of the celomic cavity may enable enhanced intrauterine stem-cell survival and greater success in achieving a functional allograft/xenograft. J. Santolaya-Forgas*, J. Deleon-Luis , I. Galan , * Division of Maternal Fetal Medicine, Department of Obstetrics and Gynecology, Brigham and Women's Hospital, 75 Francis St., Boston, MA 02115, USA, Amarillo Women's Health Research Institute, Texas Tech University Health Science Center, Amarillo, TX, USA" @default.
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• W2003641451 date "2006-07-21" @default.
• W2003641451 modified "2023-10-18" @default.
• W2003641451 title "Can extra-embryonic celomic fluid be partially replaced with stem-cell culture medium?" @default.
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• W2003641451 doi "https://doi.org/10.1002/uog.2848" @default.
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