High-Throughput Screening of Senescence Markers in Hematopoietic Stem Cells Derived from Induced Pluripotent Stem Cells
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.
Key wordsInduced pluripotent stem cells Hematopoiesis Hematopoietic stem cells Cellular senescence Telomere length
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.
- 2.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.8939CrossRefPubMedPubMedCentralGoogle Scholar
- 3.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.2013.07.03CrossRefPubMedPubMedCentralGoogle Scholar
- 5.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
- 9.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.0022261CrossRefPubMedPubMedCentralGoogle Scholar