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Series GSE132204 Query DataSets for GSE132204
Status Public on Jan 31, 2020
Title Phenotypic and functional characterization of corneal endothelial cells during in vitro expansion
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary The advent of cell culture-based methods for the establishment and expansion of human corneal endothelial cells (CEnC) has provided an available source of transplantable corneal endothelium, with a significant potential to challenge the one donor-one recipient paradigm. However, concerns over cell state identity remain, and a comprehensive characterization of the cultured CEnC has not been performed. To this end, we compared two established CEnC culture methods by assessing cell function and characterizing the transcriptomic changes that occur during in vitro expansion. In confluent monolayers, low mitogenic culture conditions preserved corneal endothelial cell state identity better than culture in high mitogenic conditions. Expansion by continuous passaging induced replicative cell senescence. Transcriptomic analysis of the senescent phenotype identified a cell senescence signature distinct for CEnC. We identified activation of both classic and new cell signaling pathways that may be targeted to prevent senescence, a significant barrier to realizing the potential clinical utility of in vitro expansion.
Overall design Seven independent primary CEnC cultures were established from donor corneas. For each culture, half the cells were prepared and isolated using Tryp-LN-F99 protocol and the other half of cells were isolated and prepared using ColA-ColIV-M4M5. Five passages (P0-P4) were established and total RNA was collected from 70 samples.
Contributor(s) Frausto RF, Swamy VS, Morselli M, Pellegrini M, Aldave AJ
Citation(s) 32366916
Submission date Jun 05, 2019
Last update date May 12, 2020
Contact name Anthony J. Aldave
Organization name Stein Eye Institute, UCLA
Department Ophthalmology
Lab Cornea Genetics Laboratory
Street address 200 Stein Plaza DSERC 3-143
City Los Angeles
State/province CA
ZIP/Postal code 90095
Country USA
Platforms (1)
GPL21290 Illumina HiSeq 3000 (Homo sapiens)
Samples (70)
GSM3853920 F99_P0_201
GSM3853921 F99_P0_277
GSM3853922 F99_P0_287
BioProject PRJNA546503
SRA SRP200456

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GSE132204_All_Processed_Data.txt.gz 6.0 Mb (ftp)(http) TXT
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