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Published on: December 16, 2021
Pancreatic-Like Cells Derived From Mouse Embryonic Stem Cells Are Regulated by Pdx1 Involving the Notch Pathway
Wa Zhong1, Yu Lai1, Zhong-Sheng Xia1
1From the Department of Gastroenterology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou.
Insights
Embryonic stem cells (ESCs) engineered to express Pdx1 successfully differentiated into pancreatic precursor cells. This advancement in Pdx1 expression is crucial for pancreas repair and cell transplantation therapies.
Area of Science:
- Stem Cell Biology
- Developmental Biology
- Regenerative Medicine
Background:
- Embryonic stem cells (ESCs) offer potential for pancreas repair.
- Expression of pancreatic duodenal homeobox 1 (Pdx1) in definitive endoderm (DE) cells is essential for pancreatic differentiation.
- A reliable model is needed to generate sufficient Pdx1-expressing DE cells for transplantation.
Purpose of the Study:
- To investigate the role of Pdx1 in regulating ESC differentiation into pancreatic cells.
- To establish a model for generating Pdx1-expressing DE cells for therapeutic applications.
Main Methods:
- A Tet-On inducible lentiviral vector system was used to express Pdx1 in mouse ESCs (ES-E14TG2a).
- ESCs were divided into control, mock vector, and Pdx1-expressing groups.
- Cells were co-cultured with sorted DE cells (CXCR-4+), and Pdx1 expression was induced by doxycycline.
- Markers of pancreatic differentiation and the Notch pathway were analyzed.
Main Results:
- Pdx1-expressing ESCs showed significantly increased expression of pancreatic markers (Ptf1a, CK19, amylase, Neuro-D1, Pax6, insulin) at various time points.
- Expression of Notch pathway components (Notch1, Notch2, Hes1, Hes5) was elevated initially and then declined.
- These findings indicate successful differentiation into pancreatic-like cells.
Conclusions:
- Pdx1-expressing ESCs can differentiate into pancreatic-like cells.
- The Notch pathway plays a role in this differentiation process.
- This model holds promise for pancreas repair and cell transplantation therapies.
Objectives:
Embryonic stem cells (ESCs)-derived pancreatic precursor cells have great potential for pancreas repair. Expression of pancreatic duodenal homeobox 1 (Pdx1) in definitive endoderm (DE) cells is the premise that DE cells differentiate into pancreatic cells. To achieve the required number of Pdx1-expressing DE cells for cell transplantation therapy, a valid model must be established. Using this model, researchers investigated how Pdx1 regulates ESC differentiation into pancreatic cells.
Methods:
Tet-On inducible lentiviral vector encoding Pdx1 or mock vector was transduced into mouse ESC (ES-E14TG2a). The mouse ESCs were divided into 3 groups: control (ESC), mock vector (Pdx1 - -ESC), and vector encoding Pdx1 (Pdx1 + -ESC). All groups were separately cocultured with the DE cells sorted by immune beads containing CXCR-4 + (C-X-C chemokine receptor type-4) antibody. Doxycycline induced the expression of Pdx1 on the Pdx1 + -ESC cells. The markers of cell differentiation and Notch pathway were examined.
Results:
Significantly increased expression levels of Ptf1a, CK19, and amylase on day (d) 3 and d7, Neuro-D1 on d10 and d14, Pax6 and insulin on d14, as well as Notch1, Notch2, Hes1, and Hes5 on d3 and thereafter declined on d14 were observed in Pdx1 + -ESC group.
Conclusions:
Pdx1 + -ESC could differentiate into pancreatic-like cells with involvement of the Notch pathway.
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