Establishment of immortalized multipotent hematopoietic progenitor cell lines by retroviral-mediated gene transfer of β-catenin

Templin, Christian; Kotlarz, Daniel; Rathinam, Chozhavendan; Rudolph, Cornelia; Schätzlein, Sonja; Ramireddy, Kodanda; Rudolph, K. Lenhard; Schlegelberger, Brigitte; Klein, Christoph; Drexler, Helmut

doi:10.1016/j.exphem.2007.10.005

Cited by 10 publications

(22 citation statements)

References 47 publications

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“…1), but failed to express lineage markers of myeloid (CD11b, GR-1, CD11c), erythroid (TER119) and lymphoid (B220, CD3e, NK1.1, CD11c) lineages. The constitutive β-catenin expression was gradually downregulated in the HPC line during 40 weeks of culture through an unknown silencing mechanism, whereas β-catenin cDNA was stably integrated in β-catenin-HPCs at 1, 8, 16, 24 and 40 weeks after transduction [22]. In the present study, we used the HPC line after 18 months of generation and culture.…”

Section: Resultsmentioning

confidence: 99%

“…β-Catenin-transduced lin– cells survived and proliferated in a cytokine cocktail of IL-3 and SCF for more than 24 months, whereas the viability of eGFP-transduced control cells dramatically decreased after 22 days of culture [22]. β-Catenin-transduced cells stably expressed (>99%) hematopoietic stem cell markers Sca-1 and c-kit (fig.…”

Section: Resultsmentioning

confidence: 99%

“…The major finding of this study is that transaortal high-dose cell transplantation with genetically modified HPCs does not alter cardiac electrophysiology in vivo or enhance ventricular automaticity in mice after I/R. We previously described genetic modification of HPCs with β-catenin, i.e., β-catenin-HPCs, as a strategy for ex vivo expansion of HPCs providing unlimited access for repair of damaged myocardium [22]. Furthermore, we reported that β-catenin-HPC transplantation caused a reduction in MI size, enhanced angiogenesis and antiapoptotic effects, associated with improvement in cardiac function in a cell dose-dependent manner [23].…”

Section: Discussionmentioning

confidence: 99%

“…The cell line was generated and characterized as described previously [22]. Briefly, human β-catenin cDNA was cloned into the retroviral vector SFβ-91-IRES-eGFP, and recombinant VSV-G-pseudotyped retroviruses were generated in the packaging cell line Phoenix-gp.…”

Section: Methodsmentioning

confidence: 99%

“…We recently reported that genetic modification of HPCs with β-catenin (β-catenin-HPCs) provided an unlimited ex vivo expansion of multipotent progenitor cells [22]. Transplantation of β-catenin-HPCs limited infarct size, attenuated left ventricular (LV) remodeling and ameliorated LV function in a cell concentration-dependent manner [23].…”

Section: Introductionmentioning

confidence: 99%

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Arrhythmia Susceptibility in Mice after Therapy with β-Catenin-Transduced Hematopoietic Progenitor Cells after Myocardial Ischemia/Reperfusion

et al. 2009

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Background: Hematopoietic progenitor cells (HPCs) can improve cardiac function after myocardial infarction. However, occurrence of arrhythmias is a potential limitation of cell therapy. In this study, we investigated the cardiac electrophysiological properties of ex vivo expanded HPCs, generated by β-catenin gene transfer, after transcoronary delivery in a murine model of ischemia/reperfusion (I/R) injury. Methods and Results: To assess arrhythmia inducibility of ex vivo expanded HPCs, mice were subjected to I/R and assigned to sham operation (n = 8), I/R (n = 21) and HPC (n = 15) treatment. Six weeks later, mice were subjected to long-term electrocardiogram recording and in vivo transvenous electrophysiological study. After I/R, mice showed a significant prolongation of conduction and repolarization compared with sham-operated mice. There was a marked increase in ventricular ectopic activity in infarcted mice as compared with sham-operated mice. Cardiac electrophysiological parameters and ventricular ectopic activity were not altered in mice treated with HPCs in comparison with control I/R mice. Conclusion: Transcoronary delivery of genetically ex vivoexpanded HPCs did not alter the electrophysiological properties in mice after I/R. Therefore, ex vivo β-catenin-mediated HPC expansion may represent an attractive therapeutic option for cell transplantation treatment of myocardial infarction without electrophysiological side effects.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Arrhythmia Susceptibility in Mice after Therapy with β-Catenin-Transduced Hematopoietic Progenitor Cells after Myocardial Ischemia/Reperfusion

et al. 2009

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TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors

Hawley

Cantor

2008

CP Stem Cell Biology

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The ability to generate genetically engineered cell lines is of great experimental value. They provide a renewable source of material that may be suitable for biochemical analyses, chromatin immunoprecipitation assays, structure‐function studies, gene function assignment, and transcription factor target gene identification. This unit describes protocols for TLX1 (HOX11)‐mediated immortalization of murine hematopoietic progenitors derived from in vitro differentiated murine embryonic stem cells, or from primary mouse fetal liver or bone marrow. A wide variety of hematopoietic cell types have been immortalized using these procedures including erythroid, megakaryocytic, monocytic, myelocytic, and multipotential cell types. These lines are typically cytokine dependent for their survival and growth. Curr. Protoc. Stem Cell Biol. 7:1F.7.1‐1F.7.19. © 2008 by John Wiley & Sons, Inc.

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Secreted proteome of the murine multipotent hematopoietic progenitor cell line DKmix

Luecke

Templin

Muetzelburg

et al. 2010

Rapid Comm Mass Spectrometry

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Administration of the multipotent hematopoietic progenitor cell (HPC) line DKmix improved cardiac function after myocardial infarction and accelerated dermal wound healing due to paracrine mechanisms. The aim of this study was to analyse the secreted proteins of DKmix cells in order to identify the responsible paracrine factors and assess their relevance to the wide spectrum of therapeutic effects. A mass spectrometry (MS)-based approach was used to identify secreted proteins of DKmix cells. Serum free culture supernatants of DKmix-conditioned medium were collected and the proteins present were separated, digested by trypsin and the resulting peptides were then analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) MS. Overall 95 different proteins were identified. Among them, secretory proteins galectin-3 and gelsolin were identified. These proteins are known to stimulate cell migration and influence wound healing and cardiac remodelling. The remaining proteins originate from intracellular compartments like cytoplasm (69%), nucleus (12%), mitochondria (4%), and cytoplasmic membrane (3%) indicating permeable or leaky DKmix cells in the conditioned medium. Additionally, a sandwich immunoassay was used to detect and quantify cytokines and chemokines. Interleukin-6 (IL-6), interleukin-13 (IL-13), monocyte-chemoattractant protein-1 (MCP-1), monocyte-chemoattractant protein-3 (MCP-3), monocyte-chemoattractant protein-1alpha (MIP-1alpha) and monocyte-chemoattractant protein-1beta (MIP-1beta) were detected in low concentrations. This study identified a subset of proteins present in the DKmix-conditioned medium that act as paracrine modulators of tissue repair. Moreover, it suggests that DKmix-derived conditioned medium might have therapeutic potency by promoting tissue regeneration.

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Establishment of immortalized multipotent hematopoietic progenitor cell lines by retroviral-mediated gene transfer of β-catenin

Cited by 10 publications

References 47 publications

Arrhythmia Susceptibility in Mice after Therapy with β-Catenin-Transduced Hematopoietic Progenitor Cells after Myocardial Ischemia/Reperfusion

Arrhythmia Susceptibility in Mice after Therapy with β-Catenin-Transduced Hematopoietic Progenitor Cells after Myocardial Ischemia/Reperfusion

TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors

Secreted proteome of the murine multipotent hematopoietic progenitor cell line DKmix

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