Derivation of human salivary epithelial progenitors from pluripotent stem cells
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA813599 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR18264635_1.fastq.gz | Fastqsanger.gz | |||
| SRR18264635_2.fastq.gz | Fastqsanger.gz | |||
| SRR18264636_1.fastq.gz | Fastqsanger.gz | |||
| SRR18264636_2.fastq.gz | Fastqsanger.gz |
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Project description:We described, for the first time, a robust protocol for direct differentiation of hPSCs into SGEP by mimicking the activity of retinoic acid and WNT signaling. These hPSC-derived SGEP expressed SOX9, KRT5 and KRT19, which were the important progenitor markers of developing salivary glands. The CD24 and α-SMA positive cells that are capable to restore the function of injured salivary glands were also present in SGEP cultures. Importantly, RNA-seq was performed to examine the similarity of SGEP to human fetal and mature salivary glands. The result revealed the SGEP resemble the transcript profile of human fetal submandibular glands. Therefore, we successfully induced hPSCs into SGEP and demonstrated that these SGEP potentially serves as models for studying mechanism underling salivary development, and as cell sources for clinical use.
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Project description:The differentiation of pluripotent stem cells has been broadly used in studying the disease mechanism and development process. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, to investigate whether the hPSCs-derived SGEPs are capable of modeling the characteristics of cystic fibrosis (CF), a disease impact SG function caused by mutation of cystic fibrosis transmembrane conductance regulator (CFTR) gene, the CFTR knockout hPSCs were differentiated into CF-SGEPs using the same protocol. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16-day of differentiation, the protein expression of CFTR was also decreased in SGEPs generated from CFTR knockout hPSCs. RNA-sequencing shows that multiple genes modulating SG development, function and were down-regulated and positive regulators of inflammation were up-regulated in CF-SGEPs, which was correlated with salivary phenotype in CF patients. These results demonstrated that suppression of CFTR disrupted the differentiation of hPSCs-derived SGEPs, which modeled the SG development of patients with CF. In summary, this study not only provided the proof that the hPSCs-derived SGEPs could serve as manipulatable and easily accessible models to study the SG developmental diseases, but also provided new avenues for the study of the CF mechanism.
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Project description:The derivation of salivary gland (SG) progenitors from pluripotent stem cells (PSCs) presents significant potential for developmental biology and regenerative medicine. However, the existing protocols for inducing SG include limited factors, making it challenging to mimic the in vivo microenvironment of embryonic SGs. We reported a cocktail factor approach to enhance the differentiation of mouse embryonic stem cell (mESC)-derived oral epithelium (OE) into SG progenitors through a three-dimensional co-culture method. Upon confirming that the embryonic SG can promote the differentiation of mESC-derived OE, we performed RNA sequence analysis to identify factors involved in the differentiation of SG progenitors. Our findings highlight several efficient pathways related to SG development, with frequent appearances of four factors: IFN-γ, TGF-β2, EGF, and IGF-1. The combined treatment using these cocktail factors increased the expression of key SG progenitor markers, including Sox9, Sox10, Krt5, and Krt14. However, absence of any one of these cocktail factors did not facilitate differentiation. Notably, aggregates treated with the cocktail factor formed SG epithelial-like structures and pre-bud-like structures on the surface. In conclusion, this study offers a novel approach to developing a differentiation protocol that closely mimics the in vivo microenvironment of embryonic SGs. This provides a foundation for generating PSC-derived SG organoids with near-physiological cell behaviors and structures.
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