Differential cytotoxic effects of mono-(2-ethylhexyl) phthalate on blastomere-derived embryonic stem cells and differentiating neurons

Chun Kyu Lim, Suel Kee Kim, Duck Sung Ko, Jea Won Cho, Jin Hyun Jun, Su Yeon An, Jung Ho Han, Jong-Hoon Kim, Yong Dal Yoon

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Potential applications of embryonic stem (ES) cells are not limited to regenerative medicine but can also include in vitro screening of various toxicants. In this study, we established mouse ES cell lines from isolated blastomeres of two-cell stage embryos and examined their potential use as an in vitro system for the study of developmental toxicity. Two ES cell lines were established from 69 blastomere-derived blastocysts (2.9%). The blastomere-derived ES (bm-ES) cells were treated with mono-(2-ethylhexyl) phthalate (MEHP) in an undifferentiated state or after directed differentiation into early neural cell types. We observed significantly decreased cell viability when undifferentiated bm-ES cells were exposed to a high dose of MEHP (1000 μM). The cytotoxic effects of MEHP were accompanied by increased DNA fragmentation, nuclear condensation, and activation of Caspase-3, which are biochemical and morphological features of apoptosis. Compared to undifferentiated bm-ES cells, considerably lower doses of MEHP (50 and 100 μM) were sufficient to induce cell death in early neurons differentiated from bm-ES cells. At the lower doses, the number of neural cells positive for the active form of Caspase-3 was greater than that for undifferentiated bm-ES cells. Thus, our data indicate that differentiating neurons are more sensitive to MEHP than undifferentiated ES cells, and that undifferentiated ES cells may have more efficient defense systems against cytotoxic stresses. These findings might contribute to the development of a new predictive screening method for assessment of hazards for developmental toxicity.

Original languageEnglish
Pages (from-to)145-154
Number of pages10
JournalToxicology
Volume264
Issue number3
DOIs
Publication statusPublished - 2009 Oct 29

Fingerprint

Blastomeres
Embryonic Stem Cells
Stem cells
Neurons
Caspase 3
Toxicity
Screening
Cell Line
Cell death
Regenerative Medicine
Blastocyst
DNA Fragmentation
mono-(2-ethylhexyl)phthalate
Condensation
Hazards
Chemical activation
Cells
Cell Survival
Apoptosis
Cell Death

Keywords

  • Apoptosis
  • Blastomere
  • Cytotoxicity
  • Embryonic stem cell
  • Mono-(2-ethylhexyl) phthalate
  • Neural progenitor cell

ASJC Scopus subject areas

  • Toxicology

Cite this

Differential cytotoxic effects of mono-(2-ethylhexyl) phthalate on blastomere-derived embryonic stem cells and differentiating neurons. / Lim, Chun Kyu; Kim, Suel Kee; Ko, Duck Sung; Cho, Jea Won; Jun, Jin Hyun; An, Su Yeon; Han, Jung Ho; Kim, Jong-Hoon; Yoon, Yong Dal.

In: Toxicology, Vol. 264, No. 3, 29.10.2009, p. 145-154.

Research output: Contribution to journalArticle

Lim, Chun Kyu ; Kim, Suel Kee ; Ko, Duck Sung ; Cho, Jea Won ; Jun, Jin Hyun ; An, Su Yeon ; Han, Jung Ho ; Kim, Jong-Hoon ; Yoon, Yong Dal. / Differential cytotoxic effects of mono-(2-ethylhexyl) phthalate on blastomere-derived embryonic stem cells and differentiating neurons. In: Toxicology. 2009 ; Vol. 264, No. 3. pp. 145-154.
@article{147c658cabb34f8c9be70354a10eb3a7,
title = "Differential cytotoxic effects of mono-(2-ethylhexyl) phthalate on blastomere-derived embryonic stem cells and differentiating neurons",
abstract = "Potential applications of embryonic stem (ES) cells are not limited to regenerative medicine but can also include in vitro screening of various toxicants. In this study, we established mouse ES cell lines from isolated blastomeres of two-cell stage embryos and examined their potential use as an in vitro system for the study of developmental toxicity. Two ES cell lines were established from 69 blastomere-derived blastocysts (2.9{\%}). The blastomere-derived ES (bm-ES) cells were treated with mono-(2-ethylhexyl) phthalate (MEHP) in an undifferentiated state or after directed differentiation into early neural cell types. We observed significantly decreased cell viability when undifferentiated bm-ES cells were exposed to a high dose of MEHP (1000 μM). The cytotoxic effects of MEHP were accompanied by increased DNA fragmentation, nuclear condensation, and activation of Caspase-3, which are biochemical and morphological features of apoptosis. Compared to undifferentiated bm-ES cells, considerably lower doses of MEHP (50 and 100 μM) were sufficient to induce cell death in early neurons differentiated from bm-ES cells. At the lower doses, the number of neural cells positive for the active form of Caspase-3 was greater than that for undifferentiated bm-ES cells. Thus, our data indicate that differentiating neurons are more sensitive to MEHP than undifferentiated ES cells, and that undifferentiated ES cells may have more efficient defense systems against cytotoxic stresses. These findings might contribute to the development of a new predictive screening method for assessment of hazards for developmental toxicity.",
keywords = "Apoptosis, Blastomere, Cytotoxicity, Embryonic stem cell, Mono-(2-ethylhexyl) phthalate, Neural progenitor cell",
author = "Lim, {Chun Kyu} and Kim, {Suel Kee} and Ko, {Duck Sung} and Cho, {Jea Won} and Jun, {Jin Hyun} and An, {Su Yeon} and Han, {Jung Ho} and Jong-Hoon Kim and Yoon, {Yong Dal}",
year = "2009",
month = "10",
day = "29",
doi = "10.1016/j.tox.2009.08.015",
language = "English",
volume = "264",
pages = "145--154",
journal = "Toxicology",
issn = "0300-483X",
publisher = "Elsevier Ireland Ltd",
number = "3",

}

TY - JOUR

T1 - Differential cytotoxic effects of mono-(2-ethylhexyl) phthalate on blastomere-derived embryonic stem cells and differentiating neurons

AU - Lim, Chun Kyu

AU - Kim, Suel Kee

AU - Ko, Duck Sung

AU - Cho, Jea Won

AU - Jun, Jin Hyun

AU - An, Su Yeon

AU - Han, Jung Ho

AU - Kim, Jong-Hoon

AU - Yoon, Yong Dal

PY - 2009/10/29

Y1 - 2009/10/29

N2 - Potential applications of embryonic stem (ES) cells are not limited to regenerative medicine but can also include in vitro screening of various toxicants. In this study, we established mouse ES cell lines from isolated blastomeres of two-cell stage embryos and examined their potential use as an in vitro system for the study of developmental toxicity. Two ES cell lines were established from 69 blastomere-derived blastocysts (2.9%). The blastomere-derived ES (bm-ES) cells were treated with mono-(2-ethylhexyl) phthalate (MEHP) in an undifferentiated state or after directed differentiation into early neural cell types. We observed significantly decreased cell viability when undifferentiated bm-ES cells were exposed to a high dose of MEHP (1000 μM). The cytotoxic effects of MEHP were accompanied by increased DNA fragmentation, nuclear condensation, and activation of Caspase-3, which are biochemical and morphological features of apoptosis. Compared to undifferentiated bm-ES cells, considerably lower doses of MEHP (50 and 100 μM) were sufficient to induce cell death in early neurons differentiated from bm-ES cells. At the lower doses, the number of neural cells positive for the active form of Caspase-3 was greater than that for undifferentiated bm-ES cells. Thus, our data indicate that differentiating neurons are more sensitive to MEHP than undifferentiated ES cells, and that undifferentiated ES cells may have more efficient defense systems against cytotoxic stresses. These findings might contribute to the development of a new predictive screening method for assessment of hazards for developmental toxicity.

AB - Potential applications of embryonic stem (ES) cells are not limited to regenerative medicine but can also include in vitro screening of various toxicants. In this study, we established mouse ES cell lines from isolated blastomeres of two-cell stage embryos and examined their potential use as an in vitro system for the study of developmental toxicity. Two ES cell lines were established from 69 blastomere-derived blastocysts (2.9%). The blastomere-derived ES (bm-ES) cells were treated with mono-(2-ethylhexyl) phthalate (MEHP) in an undifferentiated state or after directed differentiation into early neural cell types. We observed significantly decreased cell viability when undifferentiated bm-ES cells were exposed to a high dose of MEHP (1000 μM). The cytotoxic effects of MEHP were accompanied by increased DNA fragmentation, nuclear condensation, and activation of Caspase-3, which are biochemical and morphological features of apoptosis. Compared to undifferentiated bm-ES cells, considerably lower doses of MEHP (50 and 100 μM) were sufficient to induce cell death in early neurons differentiated from bm-ES cells. At the lower doses, the number of neural cells positive for the active form of Caspase-3 was greater than that for undifferentiated bm-ES cells. Thus, our data indicate that differentiating neurons are more sensitive to MEHP than undifferentiated ES cells, and that undifferentiated ES cells may have more efficient defense systems against cytotoxic stresses. These findings might contribute to the development of a new predictive screening method for assessment of hazards for developmental toxicity.

KW - Apoptosis

KW - Blastomere

KW - Cytotoxicity

KW - Embryonic stem cell

KW - Mono-(2-ethylhexyl) phthalate

KW - Neural progenitor cell

UR - http://www.scopus.com/inward/record.url?scp=70349217898&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70349217898&partnerID=8YFLogxK

U2 - 10.1016/j.tox.2009.08.015

DO - 10.1016/j.tox.2009.08.015

M3 - Article

C2 - 19720108

AN - SCOPUS:70349217898

VL - 264

SP - 145

EP - 154

JO - Toxicology

JF - Toxicology

SN - 0300-483X

IS - 3

ER -