title: Establishing a murine xenograft-model for long-term analysis of factors inducing chromosomal instability in myelodysplastic syndrome: Pitfalls and successes.
abstract: Myelodysplastic syndromes (MDS) are difficult to culture long-term showing the need of a model to study the fate of cells with MDS-abnormalities associated with chromosomal instability (CIN). This approach to establish a xenograft model transplanting human hematopoietic stem cells (HSC) with different independent lentivirally-mediated MDS-related modifications into immunodeficient mice is a long-lasting and tedious experiment with many parameters and every positive as well as non-functioning intermediate step will help the research community. As the establishment of appropriate xenograft models is increasing worldwide we aim to share our experiences to contribute toward minimizing loss of mice and following the "right" approach. Here, modified HSCs were intrafemorally transplanted into NSG and/or NSGS mice: (1) RPS14-haploinsufficiency, (2) TP53-deficiency, (3) TP53 hotspot mutations (R248W, R175H, R273H, R249S). Engraftment was achieved and cytogenetic analyses showed human cells with normal karyotypes. However, in all experiments with NSG mice, mainly control cells or GFP-negative cells were engrafted, not allowing observation of modified HSCs. In NSGS mice, engraftment rate was higher, but mice developed graft-versus-host disease. In summary, engraftment of HSCs is promising and could be used to analyze the induction of CIN. However, the analysis of modified HSCs is limited and further experiments are required to improve this model.
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Azam
authors.initials: A
authors.lastname: Salari
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Thomay
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Himmler
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Vajen
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Schienke
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Maike
authors.initials: M
authors.lastname: Hagedorn
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Ebersold
authors.affiliation: Institute of Pathology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Hans-Heinrich
authors.initials: HH
authors.lastname: Kreipe
authors.affiliation: Institute of Molecular Medicine. Goethe University Frankfurt am Main, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Krüger
authors.affiliation: Institute of Experimental Hematology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Schambach
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Schlegelberger
authors.affiliation: Institute of Human Genetics, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: goehring.gudrun@MH-Hannover.de.
authors.forename: Gudrun
authors.initials: G
authors.lastname: Göhring
mesh_terms: D000818:Animals; D043171:Chromosomal Instability; D004195:Disease Models, Animal; D055785:Gene Knockdown Techniques; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006801:Humans; D051379:Mice; D009154:Mutation; D009190:Myelodysplastic Syndromes / Q000235:genetics* / Q000473:pathology; D014183:Transplantation, Heterologous / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: D016159:Tumor Suppressor Protein p53
keywords: MDS; Mouse model; chromosomal instability; leukemia; transplantation