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Contribution of Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells to Chimeras through Injection and Coculture of Embryos.

Guo J, Wu B, Li S, Bao S, Zhao L, Hu S, Sun W, Su J, Dai Y, Li X - Stem Cells Int (2014)

Bottom Line: Blastocyst injection and morula aggregation are commonly used to evaluate stem cell pluripotency based on chimeric contribution of the stem cells.Although a significantly higher chimera rate resulted from blastocyst injection, the highest germline contribution resulted from injection of 8-cell embryos with embryonic stem cells.The fully agouti colored chimeras were generated from both injection and coculture of 8-cell embryos with embryonic stem cells.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Hohhot 010021, China ; Inner Mongolia Saikexing Reproductive Biotechnology Co., Ltd., Helingeer 011517, China.

ABSTRACT
Blastocyst injection and morula aggregation are commonly used to evaluate stem cell pluripotency based on chimeric contribution of the stem cells. To assess the protocols for generating chimeras from stem cells, 8-cell mouse embryos were either injected or cocultured with mouse embryonic stem cells and induced pluripotent stem cells, respectively. Although a significantly higher chimera rate resulted from blastocyst injection, the highest germline contribution resulted from injection of 8-cell embryos with embryonic stem cells. The fully agouti colored chimeras were generated from both injection and coculture of 8-cell embryos with embryonic stem cells. Additionally, microsatellite DNA screening showed that the fully agouti colored chimeras were fully embryonic stem cell derived mice. Unlike embryonic stem cells, the mouse chimeras were only generated from injection of 8-cell embryos with induced pluripotent stem cells and none of these showed germline transmission. The results indicated that injection of 8-cell embryos is the most efficient method for assessing stem cell pluripotency and generating induced pluripotent stem cell chimeras, embryonic stem cell chimeras with germline transmission, and fully mouse embryonic stem cell derived mice.

No MeSH data available.


Related in: MedlinePlus

Generation of mouse chimeras with iPSCs iPS. Cells were cultured on day 3 after thawing (a). Chimeric mice generated by injection of 8-cell embryos with iPSCs (b). The black arrow shows the chimeric mouse with the colored coat.
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fig4: Generation of mouse chimeras with iPSCs iPS. Cells were cultured on day 3 after thawing (a). Chimeric mice generated by injection of 8-cell embryos with iPSCs (b). The black arrow shows the chimeric mouse with the colored coat.

Mentions: Chimeric pups (13.8%, 8/58) were produced only by injection of iPSCs at passage 10–12 in 8-cell embryos (Figure 4), although nonchimeric pups were produced in the 8-cell coculture and blastocyst injection groups (Table 3). Most iPSC chimeras were fertile when mated with female KM mice, although no germline transmitting chimeras were produced.


Contribution of Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells to Chimeras through Injection and Coculture of Embryos.

Guo J, Wu B, Li S, Bao S, Zhao L, Hu S, Sun W, Su J, Dai Y, Li X - Stem Cells Int (2014)

Generation of mouse chimeras with iPSCs iPS. Cells were cultured on day 3 after thawing (a). Chimeric mice generated by injection of 8-cell embryos with iPSCs (b). The black arrow shows the chimeric mouse with the colored coat.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4291195&req=5

fig4: Generation of mouse chimeras with iPSCs iPS. Cells were cultured on day 3 after thawing (a). Chimeric mice generated by injection of 8-cell embryos with iPSCs (b). The black arrow shows the chimeric mouse with the colored coat.
Mentions: Chimeric pups (13.8%, 8/58) were produced only by injection of iPSCs at passage 10–12 in 8-cell embryos (Figure 4), although nonchimeric pups were produced in the 8-cell coculture and blastocyst injection groups (Table 3). Most iPSC chimeras were fertile when mated with female KM mice, although no germline transmitting chimeras were produced.

Bottom Line: Blastocyst injection and morula aggregation are commonly used to evaluate stem cell pluripotency based on chimeric contribution of the stem cells.Although a significantly higher chimera rate resulted from blastocyst injection, the highest germline contribution resulted from injection of 8-cell embryos with embryonic stem cells.The fully agouti colored chimeras were generated from both injection and coculture of 8-cell embryos with embryonic stem cells.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Hohhot 010021, China ; Inner Mongolia Saikexing Reproductive Biotechnology Co., Ltd., Helingeer 011517, China.

ABSTRACT
Blastocyst injection and morula aggregation are commonly used to evaluate stem cell pluripotency based on chimeric contribution of the stem cells. To assess the protocols for generating chimeras from stem cells, 8-cell mouse embryos were either injected or cocultured with mouse embryonic stem cells and induced pluripotent stem cells, respectively. Although a significantly higher chimera rate resulted from blastocyst injection, the highest germline contribution resulted from injection of 8-cell embryos with embryonic stem cells. The fully agouti colored chimeras were generated from both injection and coculture of 8-cell embryos with embryonic stem cells. Additionally, microsatellite DNA screening showed that the fully agouti colored chimeras were fully embryonic stem cell derived mice. Unlike embryonic stem cells, the mouse chimeras were only generated from injection of 8-cell embryos with induced pluripotent stem cells and none of these showed germline transmission. The results indicated that injection of 8-cell embryos is the most efficient method for assessing stem cell pluripotency and generating induced pluripotent stem cell chimeras, embryonic stem cell chimeras with germline transmission, and fully mouse embryonic stem cell derived mice.

No MeSH data available.


Related in: MedlinePlus