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Autologous transplantation of amniotic fluid-derived mesenchymal stem cells into sheep fetuses

Autologous transplantation of amniotic fluid-derived mesenchymal stem cells into sheep fetuses
Autologous transplantation of amniotic fluid-derived mesenchymal stem cells into sheep fetuses
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
0963-6897
1015-1031
Shaw, S.W. Steven
37e618c1-b931-4bc5-891e-fcec96daf7b5
Bollini, Sveva
2cb0a6ce-12ff-4e5c-8f0f-2773e7da0ead
Nader, Khalil Abi
acd79181-b937-4448-94ae-676c81ebf361
Gastadello, Annalisa
696ea861-d50a-43cd-bb50-b44c7c61ac49
Mehta, Vedanta
12aeb070-ddd9-4fe9-8935-311c4265cb15
Filppi, Elisa
64cce8c6-16bc-4249-a596-dfa74761a29f
Cananzi, Mara
b2e48d82-56ad-4e61-a56a-416bee05628c
Gaspar, H. Bobby
173d8e9b-8f39-4dbd-a441-4d739e5ea789
Qasim, Waseem
5ea8b32c-85fd-4654-94d1-ca57e01e6202
De Coppi, Paolo
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David, Anna L.
b1bdabf9-732b-424e-80a0-6e130752a206
Shaw, S.W. Steven
37e618c1-b931-4bc5-891e-fcec96daf7b5
Bollini, Sveva
2cb0a6ce-12ff-4e5c-8f0f-2773e7da0ead
Nader, Khalil Abi
acd79181-b937-4448-94ae-676c81ebf361
Gastadello, Annalisa
696ea861-d50a-43cd-bb50-b44c7c61ac49
Mehta, Vedanta
12aeb070-ddd9-4fe9-8935-311c4265cb15
Filppi, Elisa
64cce8c6-16bc-4249-a596-dfa74761a29f
Cananzi, Mara
b2e48d82-56ad-4e61-a56a-416bee05628c
Gaspar, H. Bobby
173d8e9b-8f39-4dbd-a441-4d739e5ea789
Qasim, Waseem
5ea8b32c-85fd-4654-94d1-ca57e01e6202
De Coppi, Paolo
b4236c8b-8700-4797-a529-d8791b59088b
David, Anna L.
b1bdabf9-732b-424e-80a0-6e130752a206

Shaw, S.W. Steven, Bollini, Sveva, Nader, Khalil Abi, Gastadello, Annalisa, Mehta, Vedanta, Filppi, Elisa, Cananzi, Mara, Gaspar, H. Bobby, Qasim, Waseem, De Coppi, Paolo and David, Anna L. (2011) Autologous transplantation of amniotic fluid-derived mesenchymal stem cells into sheep fetuses. Cell Transplantation, 20 (7), 1015-1031. (doi:10.3727/096368910X543402). (PMID:21092404)

Record type: Article

Abstract

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

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Published date: June 2011
Organisations: Faculty of Medicine

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Local EPrints ID: 382947
URI: http://eprints.soton.ac.uk/id/eprint/382947
ISSN: 0963-6897
PURE UUID: 8cd9cb16-9574-431d-9955-62d86113bcd8

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Date deposited: 05 Nov 2015 09:23
Last modified: 02 Dec 2019 20:30

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Contributors

Author: S.W. Steven Shaw
Author: Sveva Bollini
Author: Khalil Abi Nader
Author: Annalisa Gastadello
Author: Vedanta Mehta
Author: Elisa Filppi
Author: Mara Cananzi
Author: H. Bobby Gaspar
Author: Waseem Qasim
Author: Paolo De Coppi
Author: Anna L. David

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