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The Production of Pluripotent Stem Cells from Mouse Amniotic Fluid Cells Using a Transposon System
Published on: February 28, 2017
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Reprogramming of mouse amniotic fluid cells using a PiggyBac transposon system
E Bertin1, M Piccoli1, C Franzin1
1Stem Cells and Regenerative Medicine Laboratory, Fondazione Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy.
Stem Cell Research
|March 19, 2016
Summary
Researchers generated induced pluripotent stem (iPS) cells from mouse amniotic fluid (AF) cells using a non-viral PiggyBac transposon system. These iPS cells demonstrated pluripotency, differentiating into all three germ layers in vitro and in vivo.
Area of Science:
- Stem cell biology
- Reproductive biology
- Genetics
Background:
- Induced pluripotent stem (iPS) cells are crucial for regenerative medicine and disease modeling.
- Generating iPS cells typically involves viral methods, posing potential risks.
- Amniotic fluid (AF) is an accessible source for cell collection during prenatal diagnosis.
Purpose of the Study:
- To generate induced pluripotent stem (iPS) cells from mouse amniotic fluid (AF) cells.
- To utilize a non-viral, PiggyBac (PB) transposon-based system for iPS cell generation.
- To assess the pluripotency and differentiation capacity of the derived iPS cells.
Main Methods:
- Isolation of mouse amniotic fluid (AF) cells.
- Transfection of AF cells with the PiggyBac (PB) transposon system carrying reprogramming factors.
- Culture and selection of induced pluripotent stem (iPS) cell colonies.
- In vitro and in vivo differentiation assays to confirm pluripotency.
Main Results:
- Successfully generated multiple induced pluripotent stem (iPS) cell lines from mouse AF cells.
- The non-viral PiggyBac (PB) transposon system enabled efficient reprogramming.
- All derived iPS cell lines exhibited characteristic markers of pluripotency.
- Confirmed differentiation potential into derivatives of all three germ layers (ectoderm, mesoderm, endoderm) in vitro and in vivo.
Conclusions:
- Mouse amniotic fluid (AF) is a viable and accessible source for generating induced pluripotent stem (iPS) cells.
- The PiggyBac (PB) transposon system offers a safe and effective non-viral method for iPS cell generation from AF cells.
- The generated iPS cells possess stable pluripotency, making them valuable for research and potential therapeutic applications.
