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5. Determination of teratoma formation ability of MES cells : 4.8105MES cells were injected
CMTI-1) and established five lines of MEC cells (2A, 2B, 3G-NVRQS, 4G-NVRQS and
into subcutaneous skin of flank of 129/SvJ or nude mice. 57 days later, mice were
3G-CMTI-1). All MEC cells were remained undifferentiated by the supplementation of LIF in
sacrificedand checked the formation of teratoma histophathogically.
culture medium. The MEC cells were grown forming uniform colonies. All established MEC cells
Results : We established three lines of MES cells (9F-NVRQS, 10F-NVRQS and 11F-NVRQS)
expressed ALP, SSEA-1, Nanog, Oct-4 and Sox-2 but not SSEA-4 and TRA-1-60 and high level
and kept approximately 50 vials of frozen MES cells.Especially, 11F-NVRQS MES cell line was
of telomerase activity.
successfully maintained by 40 passages. All MES cells were remained undifferentiated by the
On the basis of these results, we report successful establishment of five MEC cell lines. We
supplementation of LIF in culture medium. The MES cells were grown forming uniform colonies.
expect the established MEC cell lines can promote our further studies of toxicity screening, drug
All established MES cells expressed ALP, SSEA-1, Nanog, Oct-4 and Sox-2 but not SSEA-4 and
discovery and cell or gene therapy.
TRA-1-60 and high level of telomerase activity.Teratomas were composed of dendrocytes,
The formation of embryoid bodies (EB) is the principal step for the differentiation of embryonic
neurocytes, keratinized squamous epithelial cells, glandular cells and fibroblasts which were
stem (ES) cells. An EB consists of ectodermal, mesodermal and endodermal tissues, which
originated from three embryonicgerm layers were observed at injection site of mice.
recapitulate many aspects of cell differentiation. For this reason, EB formation has been
Conclusions : On the basis of these results, we report successful establishment of three MES cell
utilizedwidely as a trigger of In vitro differentiation of ES cells. In this study, we established
lines. We predict the establishment of MES cell lines can promote the overall bioscience using ES
modified three different EB formation methods which were Hanging Drop (HD) method, Conical
technology for drug discovery, toxicity screening and cell or gene therapy.
Tube (CT) method and Low Attachment (LA) Method and made a comparative study of
Keyword
129/SvJ, Embryonic stem cell, ALP, SSEA-1, Nanog
characterization and formation efficacy of EB.
we established three different EB formation methods using NVRQS-11F ES cell lines. The
formation efficacy of EB was 100% in CT and LA method but 96% in HD method. The
Title
Establishment of Animal Stem Cells and Differentiated Cells
(2nd subtitle)
(Establishment, Characterization and maintenance of Animal Stem Cells)
diameter of EB by LA method was the biggest (average 653.8um). Percentage of EB presenting
Joon-Hyoung Cho*, Sang-Hee Jeong, Hyo-Shuk Shin, Young-Joo Lee, Eun-Joo
heart beating at HD method was 82.5% on 9 days of EB incubation while it was 70.9% at CT
Researcher
Kim, Jung-Hee Jang, Byoung-Kook Choi, Jeong-Woo Kang and Gab-Soo Chung
method and 39.6% at LA method. All formed EBs expressed NeuN, SSEA-1 and SSEA-4.
- Abstract -
On the basis of these results, we clarify successful establishment of three modified EB formation
Embryonic germ cells are kind of stem cells which are derived from primodial germ cells.
method using NVRQS-11F ES cell lines. There was no big differences in EB formation efficacy.
However, more EBs expressedheart beating at HD method. We expect the established EB
Mouse embryonic germ(EG) cells are usually isolated from genital ridge of embryonic day 8~12.5
which are undifferentiated. It is well-known fact that establishment of EG cell lines needed highly
formation method can promote further studies of toxicity screening, drug discovery and cell or
gene therapy through more efficient cell differentiation.
defined skill because it`s easy to differentiate during maintenance. So we used the genital ridge of
10.5days embryo to improve the efficacy. After dissection of genital ridge, EG cells were
Hepatocytes are involved in protein synthesis, carbohydrate and lipid metabolism and play a
central role in detoxication of various toxicants. So, maintenance of novel hepatocytes that contain
separated using anti- SSEA-1 and mcrobeaded secondary antibody. We established one line of
mouse EG cell line and mouse EG cells were remained undifferentiated by the supplementation of
unique characteristics is critical for reliable evaluation of in vitro hepatotoxicity of chemicals. This
study was performed to establish mouse hepatic progenitor cells (HPCs) derived from mouse
LIF in culture medium. The mouse EG cells were grown forming uniform colonies on the feeder
layer. Mouse EG cells expressed ALP, SSEA-1, Nanog, Oct-4 and Sox-2 but not SSEA-4 and
embryonic stem cells (ES). To identify and derive hepatic progenitor cells, here we developed a
3-step differentiation procedure from mouse ES-cells into HPCs and the confirmation methods of
TRA-1-60. According these results, we assure that our established mouse EG cell line is
undifferentiated noble one which has characteristics of EG.
its characteristics of morphological, biochemical, cell cycling and differentiation potential
The morphology of undifferentiated ES cell (NVRQS-11F) revealed tight, round and multi-layer
Embryonic stems isolated from teratocarcinoma, which was formed by transplantation of mouse
embryonic stem cells into extra-uterine site, are known as embryonic carcinoma (EC) cells. EC
clumps before differentiated culture, but ES-derived hepatic progenitor cells contained large nuclei
rd
and dark granular deposits within the cytoplasm after the 3
differentiation step. After
cells are also multipotential and have morphological, biological and immunological common with
embryonic stem (ES) cells. It can be a useful tool for the study of cell differentiation, cancer
differentiation, ES-HPCs stopped the expression of SSEA-1 and ALB. However, AFP, CK18,
CK19 and NESTIN continued to be expressed. Cell cycle anlaysis results demonstrated that
mechanism and variable biosciences. We have focused on the establishment of EC cell lines for
the development of fundamental technology relating to stem cell and acquirement of potent genetic
ES-HPCs cell cycle delayed than ES cells cell cycle. ES-derived hepatocytes were treated with
Periodic acid-Schiff solution and observed under light microscope. Differentiated mouse ES-derived
resources.
We obtained teratoma from testes and flank injection site of MES cell (NVRQS-11F and
hepatocytes revealed positive staining for glycogen. Differentiated mouse ES-derived bile duct cells
CMTI-1) and established five lines of MEC cells (2A, 2B, 3G-NVRQS, 4G-NVRQS and
into subcutaneous skin of flank of 129/SvJ or nude mice. 57 days later, mice were
3G-CMTI-1). All MEC cells were remained undifferentiated by the supplementation of LIF in
sacrificedand checked the formation of teratoma histophathogically.
culture medium. The MEC cells were grown forming uniform colonies. All established MEC cells
Results : We established three lines of MES cells (9F-NVRQS, 10F-NVRQS and 11F-NVRQS)
expressed ALP, SSEA-1, Nanog, Oct-4 and Sox-2 but not SSEA-4 and TRA-1-60 and high level
and kept approximately 50 vials of frozen MES cells.Especially, 11F-NVRQS MES cell line was
of telomerase activity.
successfully maintained by 40 passages. All MES cells were remained undifferentiated by the
On the basis of these results, we report successful establishment of five MEC cell lines. We
supplementation of LIF in culture medium. The MES cells were grown forming uniform colonies.
expect the established MEC cell lines can promote our further studies of toxicity screening, drug
All established MES cells expressed ALP, SSEA-1, Nanog, Oct-4 and Sox-2 but not SSEA-4 and
discovery and cell or gene therapy.
TRA-1-60 and high level of telomerase activity.Teratomas were composed of dendrocytes,
The formation of embryoid bodies (EB) is the principal step for the differentiation of embryonic
neurocytes, keratinized squamous epithelial cells, glandular cells and fibroblasts which were
stem (ES) cells. An EB consists of ectodermal, mesodermal and endodermal tissues, which
originated from three embryonicgerm layers were observed at injection site of mice.
recapitulate many aspects of cell differentiation. For this reason, EB formation has been
Conclusions : On the basis of these results, we report successful establishment of three MES cell
utilizedwidely as a trigger of In vitro differentiation of ES cells. In this study, we established
lines. We predict the establishment of MES cell lines can promote the overall bioscience using ES
modified three different EB formation methods which were Hanging Drop (HD) method, Conical
technology for drug discovery, toxicity screening and cell or gene therapy.
Tube (CT) method and Low Attachment (LA) Method and made a comparative study of
Keyword
129/SvJ, Embryonic stem cell, ALP, SSEA-1, Nanog
characterization and formation efficacy of EB.
we established three different EB formation methods using NVRQS-11F ES cell lines. The
formation efficacy of EB was 100% in CT and LA method but 96% in HD method. The
Title
Establishment of Animal Stem Cells and Differentiated Cells
(2nd subtitle)
(Establishment, Characterization and maintenance of Animal Stem Cells)
diameter of EB by LA method was the biggest (average 653.8um). Percentage of EB presenting
Joon-Hyoung Cho*, Sang-Hee Jeong, Hyo-Shuk Shin, Young-Joo Lee, Eun-Joo
heart beating at HD method was 82.5% on 9 days of EB incubation while it was 70.9% at CT
Researcher
Kim, Jung-Hee Jang, Byoung-Kook Choi, Jeong-Woo Kang and Gab-Soo Chung
method and 39.6% at LA method. All formed EBs expressed NeuN, SSEA-1 and SSEA-4.
- Abstract -
On the basis of these results, we clarify successful establishment of three modified EB formation
Embryonic germ cells are kind of stem cells which are derived from primodial germ cells.
method using NVRQS-11F ES cell lines. There was no big differences in EB formation efficacy.
However, more EBs expressedheart beating at HD method. We expect the established EB
Mouse embryonic germ(EG) cells are usually isolated from genital ridge of embryonic day 8~12.5
which are undifferentiated. It is well-known fact that establishment of EG cell lines needed highly
formation method can promote further studies of toxicity screening, drug discovery and cell or
gene therapy through more efficient cell differentiation.
defined skill because it`s easy to differentiate during maintenance. So we used the genital ridge of
10.5days embryo to improve the efficacy. After dissection of genital ridge, EG cells were
Hepatocytes are involved in protein synthesis, carbohydrate and lipid metabolism and play a
central role in detoxication of various toxicants. So, maintenance of novel hepatocytes that contain
separated using anti- SSEA-1 and mcrobeaded secondary antibody. We established one line of
mouse EG cell line and mouse EG cells were remained undifferentiated by the supplementation of
unique characteristics is critical for reliable evaluation of in vitro hepatotoxicity of chemicals. This
study was performed to establish mouse hepatic progenitor cells (HPCs) derived from mouse
LIF in culture medium. The mouse EG cells were grown forming uniform colonies on the feeder
layer. Mouse EG cells expressed ALP, SSEA-1, Nanog, Oct-4 and Sox-2 but not SSEA-4 and
embryonic stem cells (ES). To identify and derive hepatic progenitor cells, here we developed a
3-step differentiation procedure from mouse ES-cells into HPCs and the confirmation methods of
TRA-1-60. According these results, we assure that our established mouse EG cell line is
undifferentiated noble one which has characteristics of EG.
its characteristics of morphological, biochemical, cell cycling and differentiation potential
The morphology of undifferentiated ES cell (NVRQS-11F) revealed tight, round and multi-layer
Embryonic stems isolated from teratocarcinoma, which was formed by transplantation of mouse
embryonic stem cells into extra-uterine site, are known as embryonic carcinoma (EC) cells. EC
clumps before differentiated culture, but ES-derived hepatic progenitor cells contained large nuclei
rd
and dark granular deposits within the cytoplasm after the 3
differentiation step. After
cells are also multipotential and have morphological, biological and immunological common with
embryonic stem (ES) cells. It can be a useful tool for the study of cell differentiation, cancer
differentiation, ES-HPCs stopped the expression of SSEA-1 and ALB. However, AFP, CK18,
CK19 and NESTIN continued to be expressed. Cell cycle anlaysis results demonstrated that
mechanism and variable biosciences. We have focused on the establishment of EC cell lines for
the development of fundamental technology relating to stem cell and acquirement of potent genetic
ES-HPCs cell cycle delayed than ES cells cell cycle. ES-derived hepatocytes were treated with
Periodic acid-Schiff solution and observed under light microscope. Differentiated mouse ES-derived
resources.
We obtained teratoma from testes and flank injection site of MES cell (NVRQS-11F and
hepatocytes revealed positive staining for glycogen. Differentiated mouse ES-derived bile duct cells
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