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Technical Report

Preparation and analysis of fetal liver extracts

Bone Marrow Transplantation volume 26, pages 667–671 (2000)Cite this article

Abstract

The aim of this work is to describe the techniques that have been used for preparation and analysis of whole fetal liver extracts destined for in utero transplantation. Nine fetal livers between 12 and 17 weeks of gestation were prepared: cell counts and assessment of the hematopoietic cell viability were performed on cell suspensions. Hepatocytes represented 40 to 80% of the whole cell population. The remaining cells were constituted by hematopoietic cells (mainly erythroblasts), as well as by endothelial cells. The latter expressed CD34 on their surface, interfering with the assessment of CD34+ hematopoietic cells by flow cytometry. Direct visual morphologic control using alkaline phosphatase anti-alkaline phosphatase techniques was needed to differentiate hematopoietic from extra-hematopoietic CD34+ cells. Between 3.0 and 34.6 × 106 CD34+ viable hematopoietic cells were collected per fetal liver. Adequate differentiation of these cells into burst-forming units erythroid (BFU-E), colony-forming units granulocyte–macrophage (CFU-GM), and colony-forming units granulocyte erythroid macrophage megakaryocyte (CFU-GEMM) has been shown for each sample in clonogeneic cultures. In conclusion, fetal liver is a potential source of hematopoietic stem cells. Their numeration, based on the presence of CD34, is hampered by the expression of this antigen on other cells contained in the liver cell extract, in particular endothelial cells. Bone Marrow Transplantation (2000) 26, 667–671.

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Acknowledgements

The authors would like to thank Prof O Spertini and MS Quarroz for their kind advice and lending of the laboratory facilities.

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Authors and Affiliations

  1. Service Régional Vaudois de Transfusion Sanguine, Lausanne, France

    C Zwicky, D Gasparini, J-D Tissot & P Schneider

  2. Département de Gynécologie et d'Obstétrique, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

    S Gerber, F Forestier & P Hohlfeld

  3. Service de Médecine et de Biologie Foetales, Institut de Puériculture, Paris, France

    F Forestier

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  1. C Zwicky
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Zwicky, C., Gerber, S., Gasparini, D. et al. Preparation and analysis of fetal liver extracts. Bone Marrow Transplant 26, 667–671 (2000). https://doi.org/10.1038/sj.bmt.1702564

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