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Germ line chimera produced by transfer of cultured chick primordial germ cells
I K Chang1, A Yoshiki, M Kusakabe
1RIKEN Cell Bank, Institute of Physical and Chemical Research (RIKEN), Tsukuba Science City, Japan.
Insights
Cultured primordial germ cells (PGCs) from chick embryos can still migrate, colonize, and proliferate in recipient gonads, even after passing their normal migration stage. This finding has implications for germline development research.
Area of Science:
- Developmental Biology
- Cell Biology
- Reproductive Biology
Background:
- Primordial germ cells (PGCs) are essential for establishing the germline.
- Understanding PGC migration and colonization is crucial for reproductive biology.
- Previous studies have focused on PGCs within their normal developmental window.
Purpose of the Study:
- To investigate the migratory and colonization potential of cultured chick PGCs.
- To determine if PGCs, post-migration stage, retain their ability to colonize gonads.
- To assess the proliferation capacity of transplanted PGCs in a host embryo.
Main Methods:
- Isolation and in vitro culture of stage 27 chick PGCs.
- Labeling PGCs with PKH26 fluorescent dye.
- Intravenous injection of labeled PGCs into stage 17 recipient chick embryos.
- Analysis of PGC colonization and proliferation using fluorescent confocal microscopy.
Main Results:
- Labeled donor PGCs successfully migrated to the germinal ridges of recipient embryos.
- Transplanted PGCs exhibited significant proliferation, undergoing 3-7 divisions.
- Cultured PGCs, even beyond their typical migration stage, retained colonization and proliferation capabilities.
Conclusions:
- Chick PGCs retain migratory and colonization potential even after their normal developmental migration phase.
- In vitro culture does not abrogate the ability of PGCs to colonize embryonic gonads.
- These findings support the potential use of cultured PGCs for germline studies and interventions.
Abstract:
Intrinsic primordial germ cells (PGCs) from stage 27 (5-day-old) chick embryonic germinal ridges were cultured in vitro for a further 5 days, and shown to proliferate on stroma cells derived from the germinal ridge. To determine whether these cultured PGCs could colonize and contribute to the germ-line, PGCs were isolated by gentle pipetting, labeled with PKH26 fluorescent dye and injected into the blood stream of stage 17 (2.5-day-old) chick embryos. The recipient embryos were incubated until they reached stage 28. Thin sections of these embryos were analysed by fluorescent confocal laser microscopy. These analyses showed that the labeled donor PGCs had migrated into the germinal ridges of the recipient embryos, and transplanted PGCs had undergone at least 3-7 divisions. These results suggest that PGCs that had passed far beyond the migration stage in vivo were still able to migrate, colonize and proliferate in recipient chick embryonic gonads.