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Robust detection of undifferentiated iPSC among differentiated cells.
- Source :
-
Scientific reports [Sci Rep] 2020 Jun 24; Vol. 10 (1), pp. 10293. Date of Electronic Publication: 2020 Jun 24. - Publication Year :
- 2020
-
Abstract
- Recent progress in human induced pluripotent stem cells (iPSC) technologies suggest that iPSC application in regenerative medicine is a closer reality. Numerous challenges prevent iPSC application in the development of numerous tissues and for the treatment of various diseases. A key concern in therapeutic applications is the safety of the cell products to be transplanted into patients. Here, we present novel method for detecting residual undifferentiated iPSCs amongst directed differentiated cells of all three germ lineages. Marker genes, which are expressed specifically and highly in undifferentiated iPSC, were selected from single cell RNA sequence data to perform robust and sensitive detection of residual undifferentiated cells in differentiated cell products. ESRG (Embryonic Stem Cell Related), CNMD (Chondromodulin), and SFRP2 (Secreted Frizzled Related Protein 2) were well-correlated with the actual amounts of residual undifferentiated cells and could be used to detect residual cells in a highly sensitive manner using qPCR. In addition, such markers could be used to detect residual undifferentiated cells from various differentiated cells, including hepatic cells and pancreatic cells for the endodermal lineage, endothelial cells and mesenchymal cells for the mesodermal lineage, and neural cells for the ectodermal lineage. Our method facilitates robust validation and could enhance the safety of the cell products through the exclusion of undifferentiated iPSC.
- Subjects :
- Biomarkers analysis
Cell Culture Techniques
Cell Line
Colony-Forming Units Assay
Humans
Induced Pluripotent Stem Cells transplantation
Intercellular Signaling Peptides and Proteins analysis
Intercellular Signaling Peptides and Proteins genetics
Membrane Proteins analysis
Membrane Proteins genetics
Proteins analysis
Proteins genetics
RNA, Long Noncoding
RNA-Seq
Cell Differentiation genetics
Cell Separation methods
Induced Pluripotent Stem Cells physiology
Single-Cell Analysis methods
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 32581272
- Full Text :
- https://doi.org/10.1038/s41598-020-66845-6