In vitro methods to ensure absence of residual undifferentiated human induced pluripotent stem cells intermingled in induced nephron progenitor cells

Cell therapies using human induced pluripotent stem cell (hiPSC)-derived nephron progenitor cells (NPCs) are expected to ameliorate acute kidney injury (AKI). However, using hiPSC-derived NPCs clinically is a challenge because hiPSCs themselves are tumorigenic. LIN28A, ESRG, CNMD and SFRP2 transcrip...

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Bibliographic Details
Published in:PloS one 2022-11, Vol.17 (11), p.e0275600-e0275600
Main Authors: Tsujimoto, Hiraku, Katagiri, Naoko, Ijiri, Yoshihiro, Sasaki, Ben, Kobayashi, Yoshifumi, Mima, Akira, Ryosaka, Makoto, Furuyama, Kenichiro, Kawaguchi, Yoshiya, Osafune, Kenji
Format: Article
Language:English
Subjects:
Biology and Life Sciences
Biomarkers
Cell Differentiation - genetics
Clinical trials
Datasets
Flow cytometry
Gene sequencing
Genes
Health aspects
Humans
In vitro methods and tests
In Vitro Techniques
Induced Pluripotent Stem Cells - metabolism
Kidney tubules
Kidneys
Liver
Medicine and Health Sciences
Nephrons
Neuroblastoma - metabolism
Pluripotency
Progenitor cells
Proteins
Research and Analysis Methods
RNA, Long Noncoding - metabolism
Stem cells
Transplantation
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Summary:Cell therapies using human induced pluripotent stem cell (hiPSC)-derived nephron progenitor cells (NPCs) are expected to ameliorate acute kidney injury (AKI). However, using hiPSC-derived NPCs clinically is a challenge because hiPSCs themselves are tumorigenic. LIN28A, ESRG, CNMD and SFRP2 transcripts have been used as a marker of residual hiPSCs for a variety of cell types undergoing clinical trials. In this study, by reanalyzing public databases, we found a baseline expression of LIN28A, ESRG, CNMD and SFRP2 in hiPSC-derived NPCs and several other cell types, suggesting LIN28A, ESRG, CNMD and SFRP2 are not always reliable markers for iPSC detection. As an alternative, we discovered a lncRNA marker gene, MIR302CHG, among many known and unknown iPSC markers, as highly differentially expressed between hiPSCs and NPCs, by RNA sequencing and quantitative RT-PCR (qRT-PCR) analyses. Using MIR302CHG as an hiPSC marker, we constructed two assay methods, a combination of magnetic bead-based enrichment and qRT-PCR and digital droplet PCR alone, to detect a small number of residual hiPSCs in NPC populations. The use of these in vitro assays could contribute to patient safety in treatments using hiPSC-derived cells.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0275600