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Series GSE192814 Query DataSets for GSE192814
Status Public on Oct 28, 2022
Title Direct reprogramming of human fetal retinal pigment epithelial cells into photoreceptor-like cells [scRNA-seq]
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary single cell RNAseq for the CiPCs induction
 
Overall design fRPE cells were plated on 0.1% gelatin coated plates at a density of 3 x 104 cells/cm2 and cultured in fRPE medium on Day 0 (D0). The next Day (D1), medium was changed to photoreceptor induction medium 1 (PIM1) containing DMEM/F12, 10% knockout serum replacement, 2% B27, 5 ng/mL Noggin, 5 ng/mL IGF-1, in combination with 5 small molecule factors (5F): 0.5 mM VPA, 4.8 μM CHIR99021, 2 μM Repsox, 10 μM Forskolin and 10 μM IWR1. To promote and support the formation of photoreceptors, 3 nM sonic hedgehog, 100 μM taurine, 1 μM retinoic acid (STR), 1% N2 and 5 ng/mL bFGF were added in the medium (PIM2) along with 5F from D5 to D10. For PTBP1 ASO treatment, Cells were seeded on D0 and cultured in growth medium to reach a confluency at around 70%-80% 24 hours later. On D1, in addition to medium change of PIM1 with 5F, PTBP1 ASO was transfected with Lipofectamine RNAimax (ThermoFisher Scientific). We used 3.75 μL of RNAimax, 37.5 pmol ASO in 1 ml of culture medium. 48 hours after ASO treatment, medium was replaced by fresh PIM1 containing 5F and the following process was the same as described above.
 
Contributor(s) Wang J
Citation(s) 36231108
Submission date Dec 30, 2021
Last update date Oct 29, 2022
Contact name Guoping Fan
E-mail(s) guoping.fan@gmail.com
Organization name university of california, los angeles
Department Human Genetics
Street address 695 Charles E Young Drive South
City los angeles
State/province ca
ZIP/Postal code 90095
Country USA
 
Platforms (1)
GPL20301 Illumina HiSeq 4000 (Homo sapiens)
Samples (4)
GSM5765937 frpe_ctrl
GSM5765938 frpe_3d_5f
GSM5765939 frpe_3d_5fa
This SubSeries is part of SuperSeries:
GSE192882 Direct reprogramming of human fetal retinal pigment epithelial cells into photoreceptor-like cells
Relations
BioProject PRJNA793258

Download family Format
SOFT formatted family file(s) SOFTHelp
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Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE192814_RAW.tar 314.0 Mb (http)(custom) TAR (of TAR)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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