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[Establishment and differentiation of murine EG cell lines derived from primordial germ cells]
1Shanghai Institute of Cell Biology, Academia Sinica, Shanghai 200031, China.
Insights
This study successfully established five pluripotent germline-derived embryonic stem cell lines (EG cells) from early mouse embryos. These EG cells maintain pluripotency and can differentiate into various cell types, offering potential for developmental biology research.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Genetics
Context:
- Primordial germ cells (PGCs) are the precursors to gametes.
- Establishing pluripotent cell lines is crucial for developmental studies.
- Mouse models are essential for understanding early embryonic development.
Purpose:
- To isolate and characterize pluripotent embryonic germ (EG) cell lines from mouse primordial germ cells (PGCs).
- To assess the pluripotency and differentiation potential of established EG cell lines.
- To investigate the feasibility of deriving EG cells from different embryonic time points and mouse strains.
Summary:
- Primordial germ cells (PGCs) were isolated from 8.5 to 12.5 days post coitum mouse embryos and cultured with specific growth factors.
- Five EG cell lines (EG1-5) were successfully established from 8.5 and 10.5 dpc PGCs, exhibiting undifferentiated morphology and Oct-4 expression.
- These EG cell lines maintained pluripotency through 15 passages, formed embryoid bodies, differentiated into multiple cell types in vitro, and formed teratocarcinomas in vivo, confirming their pluripotency.
Impact:
- Demonstrates the successful derivation and characterization of pluripotent EG cell lines from early mouse PGCs.
- Provides valuable tools for studying germ cell development and early embryonic differentiation.
- Highlights the potential of EG cells for regenerative medicine and disease modeling.
Abstract:
Primordial germ cells (PGC) were isolated from 8.5, 10.5, 12.5 days post coitum (dpc) embryos of F1 (Balb/c x ICR), C57BL/6J, 129/svJ, 129/sv-ter mice, and cultured on mitotically inactive MEF or STO feeder layer cells with addition of leukemia inhibitory factor, stem cell factor and basic fibroblast growth factor in cultures. PGCs formed densely packed and AKP positive colonies with pluripotential marker gene (oct-4) expression resembling undifferentiated ES cells in morphology and growth pattern. Five EG cell lines derived from PGCs were established: EG1(8.5 dpc, F1), EG2 and EG3 (8.5 dpc, C57BL/6J), EG4 (10.5 dpc, 129/svJ), EG5 (10.5 dpc, 129/sv-ter). No long term culture was obtained from 12.5 dpc PGCs of 29 embryos. All five EG cell lines cultured on feeder layer cells or in LIF containing medium still remain undifferentiated state at 15 th passage. Under appropriate conditions, EG cells formed embryoid bodies in suspension culture and multiple types of differentiated cells in monolayer culture. When these EG cells were injected in nude mice, they formed teratocacinomas containing differentiated cells such as cartilage, neural tissue and epithelium. These results show that EG1-5 cell lines derived from 8.5, 10.5 dpc embryos are pluripotential.