Abstract
The successful development and characterization of human induced pluripotent stem cells (iPSCs) provides a powerful tool to study the molecular mechanisms that control cell fate decisions and differentiation toward distinct lineages. Here we focus on the ability of donors derived iPSCs to differentiate toward hematopoietic progenitor cells and on the analysis of their telomere length. The ability to screen telomere length in individual donors is important for defining cellular senescence, which correlates with their differentiation potential toward hematopoietic lineages. We have modified iPSC culture protocol and telomere length analysis to suit for high throughput screening of telomere length in large number of individual donors. This approach can be used to demonstrate the heterogeneity or changes of telomere length and its shortening as an exclusion criterion for selection of suitable donors for future stem cell therapies.
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References
Aguado T, Gutierrez FJ, Aix E, Schneider RP, Giovinnazo G, Blasco MA, Flores I (2016) Telomere length defines the cardiomyocyte differentiation potency of mouse induced pluripotent stem cells. Stem Cells 35(2):362–373. https://doi.org/10.1002/stem.2497
Lu Y, Tan CT, Nyunt MS, Mok EW, Camous X, Kared H, Fulop T, Feng L, Ng TP, Larbi A (2016) Inflammatory and immune markers associated with physical frailty syndrome: findings from Singapore longitudinal aging studies. Oncotarget 7:28783–28795. https://doi.org/10.18632/oncotarget.8939
Shao L, Wang Y, Chang J, Luo Y, Meng A, Zhou D (2013) Hematopoietic stem cell senescence and cancer therapy-induced long-term bone marrow injury. Transl Cancer Res 2:397–411. https://doi.org/10.3978/j.issn.2218-676X.2013.07.03
Ng ES, Davis R, Stanley EG, Elefanty AG (2008) A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies. Nat Protoc 3:768–776. https://doi.org/10.1038/nprot.2008.42
Ng ES, Davis RP, Hatzistavrou T, Stanley EG and Elefanty AG (2008) Directed differentiation of human embryonic stem cells as spin embryoid bodies and a description of the hematopoietic blast colony forming assay. Curr Protoc Stem Cell Biol Chapter 1:Unit 1D 3. doi: https://doi.org/10.1002/9780470151808.sc01d03s4
Tian X, Kaufman DS (2008) Hematopoietic development of human embryonic stem cells in culture. Methods Mol Biol 430:119–133. https://doi.org/10.1007/978-1-59745-182-6_8
Choi KD, Vodyanik M, Slukvin II (2011) Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells. Nat Protoc 6:296–313. https://doi.org/10.1038/nprot.2010.184
Notaro R, Cimmino A, Tabarini D, Rotoli B, Luzzatto L (1997) In vivo telomere dynamics of human hematopoietic stem cells. Proc Natl Acad Sci U S A 94:13782–13785
Niwa A, Heike T, Umeda K, Oshima K, Kato I, Sakai H, Suemori H, Nakahata T, Saito MK (2011) A novel serum-free monolayer culture for orderly hematopoietic differentiation of human pluripotent cells via mesodermal progenitors. PLoS One 6:e22261. https://doi.org/10.1371/journal.pone.0022261
Cawthon RM (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res 30:e47
O'Callaghan NJ, Fenech M (2011) A quantitative PCR method for measuring absolute telomere length. Biol Proced Online 13:3. https://doi.org/10.1186/1480-9222-13-3
Acknowledgment
Supported by European Social Fund and European Regional Development Fund—Project MAGNET (No. CZ.02.1.01/0.0/0.0/15_003/0000492). SSJ is a recipient of Masaryk University Faculty of Medicine Stipend for doctoral studies.
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Jose, S.S., Bendickova, K., Fric, J. (2018). High-Throughput Screening of Senescence Markers in Hematopoietic Stem Cells Derived from Induced Pluripotent Stem Cells. In: Ertl, P., Rothbauer, M. (eds) Cell-Based Microarrays. Methods in Molecular Biology, vol 1771. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7792-5_10
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DOI: https://doi.org/10.1007/978-1-4939-7792-5_10
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