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Derivation of Stem Cell Lines from Mouse Preimplantation Embryos
Published on: August 20, 2017
Derivation of embryonic stem cell lines from parthenogenetically developing rat blastocysts
Masumi Hirabayashi1, Teppei Goto, Chihiro Tamura
11 Section of Mammalian Transgenesis, Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences , Aichi, Japan .
Stem Cells and Development
|September 10, 2013
Summary
Researchers successfully created the first germline-competent rat embryonic stem (ES) cells from parthenogenetic blastocysts. These novel ES cells maintain key stem cell markers and epigenetic patterns, demonstrating pluripotency and potential for future research.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Reproductive Biology
Background:
- Embryonic stem (ES) cells are crucial for developmental biology research.
- Parthenogenetic development offers a potential source for generating stem cells without genetic manipulation.
- Establishing genuine ES cells from parthenogenetically derived rat blastocysts remains a significant challenge.
Purpose of the Study:
- To establish and characterize rat embryonic stem (ES) cells derived from parthenogenetically developing blastocysts.
- To assess the pluripotency and germline potential of these novel ES cells.
Main Methods:
- Parthenogenetic rat blastocysts were generated using ionomycin and cycloheximide treatment.
- ES cell lines were established using N2B27 medium with specific inhibitors (PD0325901, CHIR99021), rat LIF, and forskolin.
- Stem cell marker gene expression (Oct-4, rNanog, Fgf-4, Rex-1) was analyzed via RT-PCR.
- Epigenetic status of imprinted genes was assessed using combined bisulfite restriction analysis.
- Germline transmission potential was evaluated through chimeric rat generation.
Main Results:
- Three alkaline phosphatase-positive rat ES cell lines were successfully established from parthenogenetic blastocysts.
- All established ES cell lines expressed key stem cell markers.
- Imprinted genes in these ES cells exhibited demethylation or hypomethylation patterns similar to control ES cells.
- One ES cell line demonstrated successful germline transmission in chimeric rats.
Conclusions:
- This study reports the first successful derivation of germline-competent embryonic stem cells from parthenogenetically developing rat blastocysts.
- These findings provide a novel resource for studying pluripotency and developmental mechanisms in rats.
- The established parthenogenetic ES cells hold significant potential for future applications in regenerative medicine and genetic research.
