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Updated: Aug 17, 2026
Dechorionation of Medaka Embryos and Cell Transplantation for the Generation of Chimeras
Published on: December 22, 2010
Mouse embryos and chimera cloned from neural cells in the postnatal cerebral cortex
Hatsune Makino1, Yukiko Yamazaki, Takahiro Hirabayashi
1Department of Physiological Sciences, Graduate University for Advanced Studies, Osaka, Japan.
Abstract:
Cloning of mice has been achieved by transferring nuclei of various types of somatic cell nuclei into enucleated oocytes. However, all attempts to produce live cloned offspring using the nuclei of neurons from adult cerebral cortex have failed. Previously we obtained cloned mice using the nuclei of neural cells collected from fetal cerebral cortex. Here, we attempted to generate cloned mice using differentiated neurons from the cerebral cortex of postnatal (day 0-4) mice. Although we were unable to obtain live cloned pups, many fetuses reached day 10.5 days of development. These fetuses showed various abnormalities such as spherical omission of the neuroepithelium, collapsed lumen of neural tube, and aberrant expressions of marker proteins of neurons. We produced chimeric mice in which some hair cells and kidney cells were originated from differentiated neurons. In chimeric fetuses, LacZ-positive donor cells were in all three germ cell layers. However, chimeras with large contribution of donor-derived cells were not obtained. These results indicate that nuclei of differentiated neurons have lost their developmental totipotency. In other words, the conventional nuclear transfer technique does not allow nuclei of differentiated neurons to undergo complete genomic reprogramming required for normal embryonic development.
Insights
Cloned mice from differentiated neuron nuclei failed to develop fully. These neuron nuclei lost developmental potential, indicating incomplete genomic reprogramming for successful cloning.
Area of Science:
- Developmental Biology
- Reproductive Biology
- Neuroscience
Background:
- Somatic cell nuclear transfer (SCNT) has produced cloned mice using various somatic cell nuclei.
- Previous success involved fetal neural cell nuclei, but adult neuron nuclei failed for live offspring.
- Differentiated neurons from postnatal mice have not been tested for SCNT.
Purpose of the Study:
- To investigate the developmental totipotency of differentiated neurons from postnatal mice using SCNT.
- To determine if these nuclei can support complete embryonic development and live offspring production.
- To analyze the reprogramming efficiency of differentiated neuron nuclei during embryonic development.
Main Methods:
- Somatic cell nuclear transfer (SCNT) using differentiated neurons from postnatal (day 0-4) mouse cerebral cortex.
- Developmental assessment of reconstructed embryos and fetuses.
- Production and analysis of chimeric mice derived from differentiated neuron nuclei.
Main Results:
- No live cloned pups were obtained; however, many fetuses developed to 10.5 days.
- Significant developmental abnormalities were observed in fetuses, including neuroepithelial defects and aberrant protein expression.
- Chimeric mice showed donor cells in all germ layers, but large donor cell contributions were not achieved, suggesting limited developmental potential.
Conclusions:
- Nuclei from differentiated postnatal mouse neurons have lost developmental totipotency.
- Conventional SCNT techniques are insufficient for complete genomic reprogramming of these nuclei.
- This highlights limitations in reprogramming terminally differentiated cells for cloning purposes.
