Robust detection of undifferentiated iPSC among differentiated cells.
Biomarkers
/ analysis
Cell Culture Techniques
Cell Differentiation
/ genetics
Cell Line
Cell Separation
/ methods
Colony-Forming Units Assay
Humans
Induced Pluripotent Stem Cells
/ physiology
Intercellular Signaling Peptides and Proteins
/ analysis
Membrane Proteins
/ analysis
Proteins
/ analysis
RNA, Long Noncoding
RNA-Seq
Single-Cell Analysis
/ methods
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
24 06 2020
24 06 2020
Historique:
received:
29
07
2019
accepted:
28
05
2020
entrez:
26
6
2020
pubmed:
26
6
2020
medline:
22
12
2020
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 32581272
doi: 10.1038/s41598-020-66845-6
pii: 10.1038/s41598-020-66845-6
pmc: PMC7314783
doi:
Substances chimiques
Biomarkers
0
ESRG lncRNA, human
0
Intercellular Signaling Peptides and Proteins
0
Membrane Proteins
0
Proteins
0
RNA, Long Noncoding
0
SFRP2 protein, human
0
CNMD protein, human
136362-10-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10293Références
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