Locomotion and scanning electron microscopic observations of primordial germ cells from the embryonic chick blood in vitro
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Insights
Chick primordial germ cells (PGCs) actively migrate in vitro, moving along the axis of feeder cells. These PGCs exhibit surface microvilli and blebs, adhering to substrates primarily at their leading edge during migration.
Area of Science:
- Developmental Biology
- Cell Biology
- Embryology
Background:
- Primordial germ cells (PGCs) are crucial for reproduction, originating early in embryonic development.
- Understanding PGC migration is essential for studying germline development and potential reproductive disorders.
Purpose of the Study:
- To investigate the in vitro locomotion dynamics of chick primordial germ cells (PGCs).
- To characterize the surface morphology and substrate adhesion patterns of migrating PGCs.
Main Methods:
- Time-lapse microcinematography (16-mm and 35-mm film) was used to observe PGC movement.
- PGCs from stage 14-16 chick embryos were cultured on mesenchymal feeder cells.
- Scanning electron microscopy was employed to examine PGC surface structures post-observation.
Main Results:
- Chick PGCs demonstrated active locomotion in vitro, with an average velocity of 26 micron/hr (max 58 micron/hr).
- PGCs displayed a directional preference, moving along the longer axis of the underlying feeder cells.
- Scanning electron microscopy revealed PGCs with numerous microvilli and cytoplasmic blebs, adhering to the substrate mainly via filopodia at the leading edge.
Conclusions:
- Chick PGCs exhibit directed migration in vitro, influenced by the substrate's cellular architecture.
- The observed surface features and adhesion mechanism suggest a specific migratory strategy for PGCs.
- This study provides insights into the cellular mechanisms governing PGC migration during early development.
Abstract:
The locomotion of chick primordial germ cells (PGCs) in vitro was observed using 16-mm time-lapse microcinematography and 35-mm time-lapse film. The PGCs isolated from circulating blood of stage 14 to 16 embryos (Hamburger and Hamilton, 1951) were cultured on a substrate of mesenchymal feeder cells obtained from the dorsal mesentery of stage 40 embryos, using modified medium 199 containing 10% fetal calf serum. The PGCs were found to move actively and to show a tendency to move along the longer axis of the underlying cells. The velocity of PGC locomotion averaged 26 micron/hr and reached 58 micron/hr as a maximum. After observation, the PGCs were processed for scanning electron microscopy. They had a considerable number of microvilli about 0.2 micron in thickness and some cytoplasmic blebs on their surfaces. It was observed that the PGCs in the migrating phase adhered to the substrate with its filopodia only at the leading edge, while a large part of the cell appeared to be apart from the substrate.
