IN UTERO TRANSPLANTATION OF HUMAN STEM CELLS USING AN ANIMAL MODEL: A COMPARISON BETWEEN TWO TECHNIQUES
DOI:
https://doi.org/10.52331/cvj.v15i1.2Keywords:
stem cells, umbilical cord blood, in utero stem cell transplantation, sheep chimerasAbstract
Objective: The purpose of this study was to assess the feasibility of in utero stem cell transplantation of human umbilical cord blood stem cells in fetal sheep and to compare two different techniques of in utero transplantation, namely ultrasound-guided in utero transplantation and in utero transplantation after midline celiotomy.
Study design: Umbilical cord blood units were collected from term deliveries, after obtaining written informed consent. Human cord blood–derived, CD34+ stem cells were injected into the peritoneal cavity of 60- to 65-day-old ovine fetuses by using 2 different techniques: ultrasound-guided transabdominal percutaneous needle puncture and midline celiotomy with the exposure of the pregnant uterus. Engraftment was determined after birth by flow cytometry with use of human-specific anti-CD 34/45 antibodies.
Results: We obtained a total of 3 chimeric lambs. Using the midline celiotomy technique the fetal loss rate was 75% and only 33,3% when using ultrasound-guided transabdominal percutaneous needle puncture technique. Engraftment of donor cells was found in all fetuses, with a mean level of 1.4% in fetal peripheral blood and 3.3% in fetal bone marrow.
Conclusion: This preliminary study indicates that in utero stem cell transplantation of human hematopoietic cord blood stem cells in fetal lambs is feasible and effective in terms of hematopoietic engraftment. We also concluded that the ultrasound-guided transabdominal percutaneous needle puncture technique is more effective than performing a midline celiotomy in terms of fetal loss rate.
References
Muench MO. In utero transplantation: baby step towards an effective therapy. Bone Marrow Transplantation 2005; 35:537–47.
Flake AW, Zanjani ED. In utero hematopoietic stem cell transplantation: ontogenic opportunities and biologic barriers. Blood 1999; 94:2179-91.
Flake AW., Zanjani ED. In utero hematopoietic stem cell transplantation. A status report. JAMA 1997, vol. 278, no. 11,932937.
Westgren M. In utero stem cell transplantation. Semin. Reprod. Med 2006; 24:348-357.
Almeida-Porada G, Porada C, Gupta N, Torabi A, Thain D, Zanjani ED. The human-sheep chimeras as a model for human stem cell mobilization and evaluation of hematopoietic graft’s potential. Exp Hematol. 2007 Oct;35(10):1594-600
Porada GA, Porada C, Zanjani ED. The fetal sheep: a unique model system for assesing the full differentiative potential of human stem cells. Yonsei Med J. 2004 Jun 30; 45 Suppl: 7-14.
Porada CD, Park P, Almeida-Porada G, et al. The sheep model of in utero gene therapy. Fetal Diagn Ther 2004;19:23–30.
Miyasaka M, Morris B. The ontogeny of the lymphoid system and immune responsiveness in sheep. Prog Vet Micorbiol Immunol 1988; 4:21–55.
Almeida-Porada G, Porada CD, Tran N, Zanjani ED. Cotransplantation of human stromal cell progenitors into preimmune fetal sheep results in early appearance of human donor cells in circulation and boosts cell levels in bone marrow at later time points after transplantation. Blood 2000;95:3620–7.
Surbek DV, Young A, Danzer E, et al. Ultrasound-guided stem cell sampling from the early ovine fetus for prenatal ex vivo gene therapy. Am J Obstet Gynecol 2002; 187:960–3.
Schoeberlein A, Holzgreve W, Dudler L, et al. In utero transplantation of autologous and allogeneic fetal liver stem cells in ovine fetuses. Am J Obstet Gynecol 2004;191:1030–6.
Bernstein J, Boyle DW, Srour EF, et al. Variation in long-term engraftment of a large consecutive series of lambs transplanted in utero with human hematopoietic cells. Biol Blood Marrow Transplant 1997; 3:247–54.
Dahm-Kähler P, Wranning C, Lundmark C, Enskog A, Mölne J, Marcickiewicz J, El-Akouri RR, McCracken J, Brännström M. Transplantation of the uterus in sheep: methodology and early reperfusion events. J Obstet Gynaecol Res. 2008 Oct;34(5):784-93.
Schoeberlein A, Holzgreve W, Dudler L, Hahn S, Surbek DV. Tissue-specific engraftment after in utero transplantation of allogenic mesenchymal stem cells into sheep fetuses. Am J Obstet Gynecol. 2005 Apr; 192(4):1044-52.
Airey JA, Almeida-Porada G, Colletti EJ, Porada CD, Chamberlain J, Movsesian M, Sutko JL, Zanjani ED. Human mesenchymal stem cells form Purkinje fibers in fetal sheep heart. Circulation. 2004 Mar 23; 109(11):1401-7. Epub 2004 Mar 15.
Oppenheim SM, Muench MO, Gutiérrez-Adán A, Moyer AL, BonDurant RH, Rowe JD, Anderson GB. Hematopoietic stem cell transplantation in utero produces sheep-goat chimeras. Blood Cells Mol Dis. 2001 Jan-Feb; 27(1):296-308.
Flake AW, Roncarolo MG, Puck JM, Almeida-Porada G, Evans MI, Johnson MP, et al. Treatment of X- linked severe combined immunodeficiency by in utero transplantation of paternal bone marrow. N Engl J Med 1996; 335:1806-10.
Westgren M, Ringden O, Eik-Nes S, Ek S, Anvret M, Brubakk AM, et al. Lack of evidence of permanent engraftment after in utero fetal stem cell transplantation in congenital hemoglobinopathies. Transplantation 1996; 61:1176-9.
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