Transcription profiling of hESCs cultured on cSAPs and flat control (TCPS) using RNAseq analysis

ABSTRACT: The biophysical cues in the fate determination of pluripotent stem cells (PSCs) have not been well-revealed. In the current study, we used a novel type of substrate called colloidal self-assembled patterns (cSAPs) as an artificial matrix for the expansion of human ESCs. Briefly, the cSAPs substrate was composed of SiO2 with 2 μm diameter and PS with 100 nm diameter within the 24-well tissue culture plates (TCPS, Falcon). H1 hESCs cultured on cSAPs and flat control (TCPS) for 2 passages (4 days/passage) were dissociated with 0.5 mM EDTA for RNAseq. Total RNA (each sample ~ 1 × 106 cells) was extracted by 1 ml TRIzol (Life Technologies) and purified according to the standard instruction. RNA Sequence analysis was conducted by BGI Tech (Shenzhen, China). The analysis of gene function was performed with the multi-group data mining system of Dr. Tom (http://report.bgi.com). Gene changes on cSAPs compare to TCPS over 2-fold were defined as the significant threshold. The result indicates that hESCs cultured on the cSAPs were highly pluripotent and induced up-regulation of Toll-like receptor, HIF-1a, and Notch and down-regulation of FAs, MAPK, JAK/STAT, and TGF-β signaling pathways. We further confirmed the RNAseq result for down-regulation of CAV1, CXCL14, PTK2, and VCL genes which are part of focal adhesion and MAPK signaling by qPCR analysis. These findings demonstrate that the complex cSAPs can generate specific biophysical cues for the pluripotency of PSCs and enhance the differentiation potential to hemopoietic cells. We further demonstrated that the cSAPs substrate can keep the pluripotency and increase the efficiency of hemopoietic differentiation of cultured PSCs in comparison with the cultured cells on the flat substrate.

INSTRUMENT(S): BGISEQ-500

ORGANISM(S): Homo sapiens

SUBMITTER: Javad Harati 

PROVIDER: E-MTAB-11123 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress