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Establishment of rat embryonic stem cells and making of chimera rats.

Ueda S, Kawamata M, Teratani T, Shimizu T, Tamai Y, Ogawa H, Hayashi K, Tsuda H, Ochiya T - PLoS ONE (2008)

Bottom Line: Here we have established two independent rES cells from Wister rat blastocysts that have undifferentiated characters such as Nanog and Oct3/4 genes expression and they have stage-specific embryonic antigen (SSEA) -1, -3, -4, and TRA-1-81 expression.The cells were successfully cultured in an undifferentiated state and can be possible over 18 passages with maintaining more than 40% of normal karyotype.Their pluripotent potential was confirmed by the differentiation into derivatives of the endoderm, mesoderm, and ectoderm.

View Article: PubMed Central - PubMed

Affiliation: Section for Studies on Metastasis, National Cancer Center Research Institute, Tokyo, Japan.

ABSTRACT
The rat is a reference animal model for physiological studies and for the analysis of multigenic human diseases such as hypertension, diabetes, neurological disorders, and cancer. The rats have long been used in extensive chemical carcinogenesis studies. Thus, the rat embryonic stem (rES) cell is an important resource for the study of disease models. Attempts to derive ES cells from various mammals, including the rat, have not succeeded. Here we have established two independent rES cells from Wister rat blastocysts that have undifferentiated characters such as Nanog and Oct3/4 genes expression and they have stage-specific embryonic antigen (SSEA) -1, -3, -4, and TRA-1-81 expression. The cells were successfully cultured in an undifferentiated state and can be possible over 18 passages with maintaining more than 40% of normal karyotype. Their pluripotent potential was confirmed by the differentiation into derivatives of the endoderm, mesoderm, and ectoderm. Most importantly, the rES cells are capable of producing chimera rats. Therefore, we established pluripotent rES cell lines that are widely used to produce genetically modified experimental rats for study of human diseases.

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Pluripotency of rat ES cells.Embryoid bodies formation (A) Ws-4-1and (B) Ws-4-2. Scale bar, 200 µm. Immunostaining confirming in vitro differentiation into all three germ layers (Ws-4-1). Expression of (C) beta-III tubulin (ectoderm), (D) cytokeratin 18 (endoderm) and (E) alpha-SMA (mesoderm). Scale bar, 50 µm. (F) Ws-9 derived tumor after subcutaneous injection into SCID-mouse. Hematoxylin and eosin stained histological sections of rES (Ws-9) derived tumor: (G) intestinal epithelium-like structure (endoderm) and (H) cartilage-like structure (mesoderm).
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pone-0002800-g002: Pluripotency of rat ES cells.Embryoid bodies formation (A) Ws-4-1and (B) Ws-4-2. Scale bar, 200 µm. Immunostaining confirming in vitro differentiation into all three germ layers (Ws-4-1). Expression of (C) beta-III tubulin (ectoderm), (D) cytokeratin 18 (endoderm) and (E) alpha-SMA (mesoderm). Scale bar, 50 µm. (F) Ws-9 derived tumor after subcutaneous injection into SCID-mouse. Hematoxylin and eosin stained histological sections of rES (Ws-9) derived tumor: (G) intestinal epithelium-like structure (endoderm) and (H) cartilage-like structure (mesoderm).

Mentions: EBs were produced by culturing rES cell aggregates in Petri dishes. Ws-4-1 and Ws-9 rES cells formed simple EBs within a few days (Figure 2A). Especially, when these cultures were maintained, simple EBs of Ws-4-2 developed cystic EBs (Figure 2B).


Establishment of rat embryonic stem cells and making of chimera rats.

Ueda S, Kawamata M, Teratani T, Shimizu T, Tamai Y, Ogawa H, Hayashi K, Tsuda H, Ochiya T - PLoS ONE (2008)

Pluripotency of rat ES cells.Embryoid bodies formation (A) Ws-4-1and (B) Ws-4-2. Scale bar, 200 µm. Immunostaining confirming in vitro differentiation into all three germ layers (Ws-4-1). Expression of (C) beta-III tubulin (ectoderm), (D) cytokeratin 18 (endoderm) and (E) alpha-SMA (mesoderm). Scale bar, 50 µm. (F) Ws-9 derived tumor after subcutaneous injection into SCID-mouse. Hematoxylin and eosin stained histological sections of rES (Ws-9) derived tumor: (G) intestinal epithelium-like structure (endoderm) and (H) cartilage-like structure (mesoderm).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2483735&req=5

pone-0002800-g002: Pluripotency of rat ES cells.Embryoid bodies formation (A) Ws-4-1and (B) Ws-4-2. Scale bar, 200 µm. Immunostaining confirming in vitro differentiation into all three germ layers (Ws-4-1). Expression of (C) beta-III tubulin (ectoderm), (D) cytokeratin 18 (endoderm) and (E) alpha-SMA (mesoderm). Scale bar, 50 µm. (F) Ws-9 derived tumor after subcutaneous injection into SCID-mouse. Hematoxylin and eosin stained histological sections of rES (Ws-9) derived tumor: (G) intestinal epithelium-like structure (endoderm) and (H) cartilage-like structure (mesoderm).
Mentions: EBs were produced by culturing rES cell aggregates in Petri dishes. Ws-4-1 and Ws-9 rES cells formed simple EBs within a few days (Figure 2A). Especially, when these cultures were maintained, simple EBs of Ws-4-2 developed cystic EBs (Figure 2B).

Bottom Line: Here we have established two independent rES cells from Wister rat blastocysts that have undifferentiated characters such as Nanog and Oct3/4 genes expression and they have stage-specific embryonic antigen (SSEA) -1, -3, -4, and TRA-1-81 expression.The cells were successfully cultured in an undifferentiated state and can be possible over 18 passages with maintaining more than 40% of normal karyotype.Their pluripotent potential was confirmed by the differentiation into derivatives of the endoderm, mesoderm, and ectoderm.

View Article: PubMed Central - PubMed

Affiliation: Section for Studies on Metastasis, National Cancer Center Research Institute, Tokyo, Japan.

ABSTRACT
The rat is a reference animal model for physiological studies and for the analysis of multigenic human diseases such as hypertension, diabetes, neurological disorders, and cancer. The rats have long been used in extensive chemical carcinogenesis studies. Thus, the rat embryonic stem (rES) cell is an important resource for the study of disease models. Attempts to derive ES cells from various mammals, including the rat, have not succeeded. Here we have established two independent rES cells from Wister rat blastocysts that have undifferentiated characters such as Nanog and Oct3/4 genes expression and they have stage-specific embryonic antigen (SSEA) -1, -3, -4, and TRA-1-81 expression. The cells were successfully cultured in an undifferentiated state and can be possible over 18 passages with maintaining more than 40% of normal karyotype. Their pluripotent potential was confirmed by the differentiation into derivatives of the endoderm, mesoderm, and ectoderm. Most importantly, the rES cells are capable of producing chimera rats. Therefore, we established pluripotent rES cell lines that are widely used to produce genetically modified experimental rats for study of human diseases.

Show MeSH
Related in: MedlinePlus