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Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
- Source :
- Stem Cell Reports, Stem Cell Reports, Vol 5, Iss 4, Pp 660-671 (2015)
- Publisher :
- The Authors. Published by Elsevier Inc.
-
Abstract
- Summary The collection sites of human primary tissue samples and the receiving laboratories, where the human induced pluripotent stem cells (hIPSCs) are derived, are often not on the same site. Thus, the stability of samples prior to derivation constrains the distance between the collection site and the receiving laboratory. To investigate sample stability, we collected blood and held it at room temperature for 5, 24, or 48 hr before isolating peripheral blood mononuclear cells (PBMCs) and reprogramming into IPSCs. Additionally, PBMC samples at 5- and 48-hr time points were frozen in liquid nitrogen for 4 months and reprogrammed into IPSCs. hIPSC lines derived from all time points were pluripotent, displayed no marked difference in chromosomal aberration rates, and differentiated into three germ layers. Reprogramming efficiency at 24- and 48-hr time points was 3- and 10-fold lower, respectively, than at 5 hr; the freeze-thaw process of PBMCs resulted in no obvious change in reprogramming efficiency.<br />Graphical Abstract<br />Highlights • hIPSCs were derived from blood samples stored for 2 days at room temperature • Blood held for 24 and 48 hr produced less IPSC colonies than at 5 hr • Increasing input cells and virus volumes rescued the drop in reprogramming efficiency • IPSCs from all three storage time points can differentiate into three germ layers<br />Agu and colleagues investigated the permissible time during which blood samples can be stored at room temperature after collection without severely compromising their ability to be successfully reprogrammed into hIPSCs. They found that IPSCs can be derived from blood stored for 2 days at room temperature. The derived IPSCs maintained genomic stability and can differentiate into three germ layers.
- Subjects :
- Resource
Cellular differentiation
Induced Pluripotent Stem Cells
Germ layer
Cell Separation
Biology
Biochemistry
Peripheral blood mononuclear cell
Cryopreservation
Genomic Instability
03 medical and health sciences
0302 clinical medicine
Genetics
Humans
Cellular Reprogramming Techniques
Induced pluripotent stem cell
lcsh:QH301-705.5
Cells, Cultured
030304 developmental biology
0303 health sciences
lcsh:R5-920
Temperature
Cell Differentiation
Cell Biology
Cellular Reprogramming
Sample stability
Cell biology
lcsh:Biology (General)
Blood Preservation
Leukocytes, Mononuclear
lcsh:Medicine (General)
Reprogramming
030217 neurology & neurosurgery
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 22136711
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Stem Cell Reports
- Accession number :
- edsair.doi.dedup.....e6876fa98f88941b981758ec4910d321
- Full Text :
- https://doi.org/10.1016/j.stemcr.2015.08.012