TY  - JOUR
A1  - Olmer, Ruth
A1  - Engels, Lena
A1  - Usman, Abdulai
A1  - Menke, Sandra
A1  - Malik, Muhammad Nasir Hayat
A1  - Pessler, Frank
A1  - Goehring, Gudrun
A1  - Bornhorst, Dorothee
A1  - Bolten, Svenja
A1  - Abdelilah-Seyfried, Salim
A1  - Scheper, Thomas
A1  - Kempf, Henning
A1  - Zweigerdt, Robert
A1  - Martin, Ulrich
T1  - Differentiation of Human Pluripotent Stem Cells into Functional Endothelial Cells in Scalable Suspension Culture
JF  - Stem Cell Reports
N2  - Endothelial cells (ECs) are involved in a variety of cellular responses. As multifunctional components of vascular structures, endothelial (progenitor) cells have been utilized in cellular therapies and are required as an important cellular component of engineered tissue constructs and in vitro disease models. Although primary ECs from different sources are readily isolated and expanded, cell quantity and quality in terms of functionality and karyotype stability is limited. ECs derived from human induced pluripotent stem cells (hiPSCs) represent an alternative and potentially superior cell source, but traditional culture approaches and 2D differentiation protocols hardly allow for production of large cell numbers. Aiming at the production of ECs, we have developed a robust approach for efficient endothelial differentiation of hiPSCs in scalable suspension culture. The established protocol results in relevant numbers of ECs for regenerative approaches and industrial applications that show in vitro proliferation capacity and a high degree of chromosomal stability.
KW  - virus infection
KW  - progenitor cells
KW  - in vitro
KW  - telomere dysfunction
KW  - cord blood
KW  - cardiomyogenic differentiation
KW  - angiogenesis
KW  - efficient
KW  - aberrations
KW  - expression
Y1  - 2017
U6  - https://doi.org/10.1016/j.stemcr.2018.03.017
SN  - 2213-6711
VL  - 10
IS  - 5
PB  - Springer
CY  - New York
ER  - 
TY  - JOUR
A1  - Moradian, Hanieh
A1  - Roch, Toralf
A1  - Lendlein, Andreas
A1  - Gossen, Manfred
T1  - mRNA transfection-induced activation of primary human monocytes and macrophages
BT  - Dependence on carrier system and nucleotide modifcation
JF  - Scientific reports
N2  - Monocytes and macrophages are key players in maintaining immune homeostasis. Identifying strategies to manipulate their functions via gene delivery is thus of great interest for immunological research and biomedical applications. We set out to establish conditions for mRNA transfection in hard-to-transfect primary human monocytes and monocyte-derived macrophages due to the great potential of gene expression from in vitro transcribed mRNA for modulating cell phenotypes. mRNA doses, nucleotide modifications, and different carriers were systematically explored in order to optimize high mRNA transfer rates while minimizing cell stress and immune activation. We selected three commercially available mRNA transfection reagents including liposome and polymer-based formulations, covering different application spectra. Our results demonstrate that liposomal reagents can particularly combine high gene transfer rates with only moderate immune cell activation. For the latter, use of specific nucleotide modifications proved essential. In addition to improving efficacy of gene transfer, our findings address discrete aspects of innate immune activation using cytokine and surface marker expression, as well as cell viability as key readouts to judge overall transfection efficiency. The impact of this study goes beyond optimizing transfection conditions for immune cells, by providing a framework for assessing new gene carrier systems for monocyte and macrophage, tailored to specific applications.
KW  - sirna transfection
KW  - mediated delivery
KW  - gene delivery
KW  - efficient
KW  - immunogenicity
KW  - lipoplexes
KW  - cells
KW  - therapeutics
KW  - polarization
KW  - pathways
Y1  - 2020
U6  - https://doi.org/10.1038/s41598-020-60506-4
SN  - 2045-2322
VL  - 10
IS  - 1
SP  - 1
EP  - 15
PB  - Springer Nature
CY  - London
ER  -