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Series GSE131065 Query DataSets for GSE131065
Status Public on Dec 31, 2019
Title bulk RNA-seq of human nucleusus pulposus from scoliosis patients
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Intervertebral disc degeneration (IDD) leads to low back pain and disability globally. Progressive loss of nucleus pulposus cells (NPCs) are associated with the onset of IDD. Cell-based therapy has been shown the promising for many diseases, including the IDD in preclinical studies. However, the limited availability of human NPCs has hurdled such application for IDD. This study aimed to define strategies to derive NPCs from human ESC/iPSC. Human ESC3, ESC9 and IMR-90-iPSC were used for a three-step protocol to direct differentiation toward mesodermal, subsequently notochordal and finally NPCs. Our results showed that notochordal-like cells (NCCs) were successfully derived from the first two-step of protocol confirmed by immunofluorescence staining of Noto, Brachyury (T) and Foxa2. RT-PCR results showed that the expression of Noto, T and Foxa2 was the highest at day 5 after differentiation. Furthermore, these NCCs can be differentiated into NPCs via the final step of the protocol. These NPCs expressed the tyrosine kinase receptor Tie2 (Tie2), disialoganglioside 2 (GD2), collagen II and aggrecan. Genome-wide transcriptomic analyses by sequencing (RNA-seq) further reveals the expression of a wide array of extra-cellular matrix (ECM) genes, up-stream regulators and pathways. Cross-comparison of our RNA-seq profiles with public NPC data confirms the differentiated products are much closer to NPCs than are ESC/iPSC. Co-culturing NPCs with Light2 cell line, a luciferase-based reporter responsive to sonic hedgehog (Shh) secreted protein, showed NPCs synthesized Shh. Transplantation of NPCs effectively attenuated the spinal injury in a puncture-induced disc degeneration (PDD) model in rat. Furthermore, we utilized CRISPR/Cas9 technology to seamlessly knock in an enhanced fluorescent protein (GFP) to the loci of the noto gene in ESC9. Our study demonstrates that NPCs could be induced from human ESC/iPSC. This study provides a cell source for the development of novel strategy for IDD diseases.
 
Overall design We sequenced four NP from three scoliosis individuals
 
Contributor(s) Chen P, Au T, Tam V, SE Cheah K
Citation(s) 32101752
Submission date May 12, 2019
Last update date Feb 29, 2020
Contact name Peikai Chen
E-mail(s) pkchen1@hku.hk
Phone 852-258315217
Organization name The University of Hong Kong
Department Faculty of Medicine
Street address Sassoon Road 5
City Pok Fu Lam
ZIP/Postal code 10000
Country Hong Kong
 
Platforms (1)
GPL18460 Illumina HiSeq 1500 (Homo sapiens)
Samples (4)
GSM3762919 sample 124
GSM3762920 sample135a
GSM3762921 sample 135b
Relations
BioProject PRJNA542614
SRA SRP198229

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

Supplementary file Size Download File type/resource
GSE131065_invivoNPC.gene.expression.tables.FPKM.txt.gz 990.1 Kb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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