Skip to main content
Top
Published in: Neurotoxicity Research 3/2017

01-04-2017 | ORIGINAL ARTICLE

Human Umbilical Cord Blood-Derived Neural Stem Cell Line as a Screening Model for Toxicity

Authors: Rajashree Patnaik, Rabindra Nath Padhy

Published in: Neurotoxicity Research | Issue 3/2017

Login to get access

Abstract

The aim was to investigate whether a human neural stem cell (NSC) line derived from human umbilical cord blood (hUCB) can be used for toxicity study. Toxicity of both neurotoxic environmental xenobiotics, methyl mercury chloride (CH3HgCl), lead acetate (CH3COOPb), and chlorpyrifos (CP), and non-neurotoxic insecticide, dichlorvos, as well as non-neurotoxic drugs, theophylline and acetaminophen were assessed. Additionally, differentiation of neuronal and glial cell lines derived from hUCB was elucidated. It was observed that CH3HgCl was more toxic to human NSCs in comparison to CH3COOPb and CP. The minimum inhibitory concentration (MIC) value against NSCs was 3, 10, and 300 mg/L, in each staining process, acridine orange/ethidium bromide (AO/EB) staining, 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay, and Hoechst staining, for CH3HgCl, CP, and CH3COOPb, respectively. CH3HgCl had the LC25 value as 10.0, 14.4, and 12.7 mg/L, by staining method mentioned in succession. CP had the LC25 value as 21.9, 23.7, and 18.4 mg/L; similarly, CH3COOPb had LC25 values, successively as 616.9, 719.2, and 890.3 mg/L. LC50 values ranged from 18.2 to 21.7 mg/L for CH3HgCl, 56.4 to 60.2 mg/L for CP, and 1000 to 1460.1 for CH3COOPb. Theophylline, acetaminophen, and dichlorvos had no impact on the viability of NSCs. This work justified that hUCB-NSC model can be used for toxicity study.
Literature
go back to reference Akande MG, Aliu YO, Ambali SF (2014) Protective effect of taurine in chlorpyrifos and lead-induced hematological alterations in Wistar rats. Toxicol Environ Saf 96:171–182CrossRef Akande MG, Aliu YO, Ambali SF (2014) Protective effect of taurine in chlorpyrifos and lead-induced hematological alterations in Wistar rats. Toxicol Environ Saf 96:171–182CrossRef
go back to reference Bellinger DC (2008) Very low lead exposures and children’s neurodevelopment. Curr Opin Pediatr 20:172–177CrossRefPubMed Bellinger DC (2008) Very low lead exposures and children’s neurodevelopment. Curr Opin Pediatr 20:172–177CrossRefPubMed
go back to reference Bishayi B, Sengupta M (2006) Synergism in immunotoxicological eff ects due to repeated combined administration of arsenic and lead in mice. Int Immunopharmacol 6:454–464CrossRefPubMed Bishayi B, Sengupta M (2006) Synergism in immunotoxicological eff ects due to repeated combined administration of arsenic and lead in mice. Int Immunopharmacol 6:454–464CrossRefPubMed
go back to reference Breier JM, Gassmann K, Kayser R, Stegemen H, Groot DD, Fritsche E, Shafer TJ (2010) Neural progenitor cells as models for high-throughput screens of developmental neurotoxicity: state of the science. Neurotoxicol Teratol 32:4–15CrossRefPubMed Breier JM, Gassmann K, Kayser R, Stegemen H, Groot DD, Fritsche E, Shafer TJ (2010) Neural progenitor cells as models for high-throughput screens of developmental neurotoxicity: state of the science. Neurotoxicol Teratol 32:4–15CrossRefPubMed
go back to reference Buzanska L, Sypecka J, Nerini-Molteni S, Compagnoni A, Hogberg HT, Torchio RD, Domanska-Janik K, Zimmer J, Coecke S (2009) A human stem cell-based model for identifying adverse effects of organic and inorganic chemicals on the developing nervous system. Stem Cells 27:2591–2601CrossRefPubMed Buzanska L, Sypecka J, Nerini-Molteni S, Compagnoni A, Hogberg HT, Torchio RD, Domanska-Janik K, Zimmer J, Coecke S (2009) A human stem cell-based model for identifying adverse effects of organic and inorganic chemicals on the developing nervous system. Stem Cells 27:2591–2601CrossRefPubMed
go back to reference Caldwell MA, He X, Wilkie N (2001) Growth factors regulate the survival and fate of cells derived from human neurospheres. Nat Biotechnol 19:475–479CrossRefPubMed Caldwell MA, He X, Wilkie N (2001) Growth factors regulate the survival and fate of cells derived from human neurospheres. Nat Biotechnol 19:475–479CrossRefPubMed
go back to reference Cocks G, Romanyuk N, Amemori T, Jendelova P, Forostyak O, Jeffries AR, Perfect L, Thuret S, Dayanithi G, Sykova E, Price J (2013) Conditionally immortalized stem cell lines from human spinal cord retain regional identity and generate functional V2a interneurons and motorneurons. Stem Cell Res Ther 4:69. doi:10.1186/scrt220 CrossRefPubMedPubMedCentral Cocks G, Romanyuk N, Amemori T, Jendelova P, Forostyak O, Jeffries AR, Perfect L, Thuret S, Dayanithi G, Sykova E, Price J (2013) Conditionally immortalized stem cell lines from human spinal cord retain regional identity and generate functional V2a interneurons and motorneurons. Stem Cell Res Ther 4:69. doi:10.​1186/​scrt220 CrossRefPubMedPubMedCentral
go back to reference De Filippis L, Lamorte G, Snyder EY, Malgaroli A, Vescovi AL (2007) A novel, immortal, and multipotent human neural stem cell line generating functional neurons and oligodendrocytes. Stem Cells 25:2312–2321CrossRefPubMed De Filippis L, Lamorte G, Snyder EY, Malgaroli A, Vescovi AL (2007) A novel, immortal, and multipotent human neural stem cell line generating functional neurons and oligodendrocytes. Stem Cells 25:2312–2321CrossRefPubMed
go back to reference Divya MS, Roshin GE, Divya TS, Rasheed VA, Santhoshkumar TR, Elizabeth KE, James J, Pillai RM (2012) Umbilical cord blood-derived mesenchymal stem cells consist of a unique population of progenitors co-expressing mesenchymal stem cell and neuronal markers capable of instantaneous neuronal differentiation. Stem Cell Res Ther 3:57. doi:10.1186/scrt148 CrossRefPubMedPubMedCentral Divya MS, Roshin GE, Divya TS, Rasheed VA, Santhoshkumar TR, Elizabeth KE, James J, Pillai RM (2012) Umbilical cord blood-derived mesenchymal stem cells consist of a unique population of progenitors co-expressing mesenchymal stem cell and neuronal markers capable of instantaneous neuronal differentiation. Stem Cell Res Ther 3:57. doi:10.​1186/​scrt148 CrossRefPubMedPubMedCentral
go back to reference Dubrez L, Savoy I, Hamman A, Solary E (1996) Pivotal role of a DEVD-sensitive step in etoposide-induced and Fas-mediated apoptotic pathways. EMBO J 15:5504–5512PubMedPubMedCentral Dubrez L, Savoy I, Hamman A, Solary E (1996) Pivotal role of a DEVD-sensitive step in etoposide-induced and Fas-mediated apoptotic pathways. EMBO J 15:5504–5512PubMedPubMedCentral
go back to reference Fu L, Zhu L, Huang Y, Lee TD, Forman SJ, Shih CC (2008) Derivation of neural stem cells from mesenchymal stem cells: evidence for a bipotential stem cell population. Stem Cells Dev 17:1109–1121CrossRefPubMedPubMedCentral Fu L, Zhu L, Huang Y, Lee TD, Forman SJ, Shih CC (2008) Derivation of neural stem cells from mesenchymal stem cells: evidence for a bipotential stem cell population. Stem Cells Dev 17:1109–1121CrossRefPubMedPubMedCentral
go back to reference Hargreaves AJ, Sachana M, Flaskos J (2011) The use of differentiating N2a and C6 cell lines for studies of organophosphate toxicity. Cell Culture Techniq 56:269–291CrossRef Hargreaves AJ, Sachana M, Flaskos J (2011) The use of differentiating N2a and C6 cell lines for studies of organophosphate toxicity. Cell Culture Techniq 56:269–291CrossRef
go back to reference Imai K, Kusakawa S, Tanoue A, Kuwagata M, Senuma M, Furuya M, Takashima H (2008) In vitro embryotoxicity testing of mercury vapour by differentiation of ES-D3 cells. Altern Animal Test Experimen 13:118–122 Imai K, Kusakawa S, Tanoue A, Kuwagata M, Senuma M, Furuya M, Takashima H (2008) In vitro embryotoxicity testing of mercury vapour by differentiation of ES-D3 cells. Altern Animal Test Experimen 13:118–122
go back to reference Huang FN, Schneider JS (2004) Effects of lead exposure on proliferation and differentiation of neural stem cells derived from different regions of embryonic rats brain. Neurotoxicology 25:1001–1012CrossRefPubMed Huang FN, Schneider JS (2004) Effects of lead exposure on proliferation and differentiation of neural stem cells derived from different regions of embryonic rats brain. Neurotoxicology 25:1001–1012CrossRefPubMed
go back to reference Klaric M, Winkler J, Vojnits K, Meganathan K, Jagtap S, Ensenat-Waser R, Hescheler J, Sachinidis A, Bremer-Hoffmann S (2013) Current status of human pluripotent stem cell based in vitro toxicity tests. Front Biosci 5:118–133CrossRef Klaric M, Winkler J, Vojnits K, Meganathan K, Jagtap S, Ensenat-Waser R, Hescheler J, Sachinidis A, Bremer-Hoffmann S (2013) Current status of human pluripotent stem cell based in vitro toxicity tests. Front Biosci 5:118–133CrossRef
go back to reference Makris SL, Raffaele K, Allen S, Bowers WJ, Hass U, Alleva E, Calamandrei G, Sheets L, Amcoff P, Delrue N, Crofton KM (2009) A retrospective performance assessment of the developmental neurotoxicity study in support of OECD test guideline 426. Environ Health Perspect 117:17–25CrossRefPubMed Makris SL, Raffaele K, Allen S, Bowers WJ, Hass U, Alleva E, Calamandrei G, Sheets L, Amcoff P, Delrue N, Crofton KM (2009) A retrospective performance assessment of the developmental neurotoxicity study in support of OECD test guideline 426. Environ Health Perspect 117:17–25CrossRefPubMed
go back to reference Marasinghe J, Yu Q, Connell D (2014) Assessment of health risk in human populations due to chlorpyrifos. Toxics 2:92–114CrossRef Marasinghe J, Yu Q, Connell D (2014) Assessment of health risk in human populations due to chlorpyrifos. Toxics 2:92–114CrossRef
go back to reference Markiewicz I, Sypecka J, Domanska-Janik K, Wyszomirski T, Lukomska B (2011) Cellular environment directs differentiation of human umbilical cord blood–derived neural stem cells in vitro. J Histochem Cytochem 59:289–301CrossRefPubMedPubMedCentral Markiewicz I, Sypecka J, Domanska-Janik K, Wyszomirski T, Lukomska B (2011) Cellular environment directs differentiation of human umbilical cord blood–derived neural stem cells in vitro. J Histochem Cytochem 59:289–301CrossRefPubMedPubMedCentral
go back to reference Mezey E, Key S, Vogelsang G, Szalayova I, Lange GD, Crain B (2003) Transplanted bone marrow generates new neurons in human brains. Proc Natl Acad Sci U S A 4:1364–1369CrossRef Mezey E, Key S, Vogelsang G, Szalayova I, Lange GD, Crain B (2003) Transplanted bone marrow generates new neurons in human brains. Proc Natl Acad Sci U S A 4:1364–1369CrossRef
go back to reference Miranda RC, Santillano DR, Camarillo C, Dohrman D (2008) Modeling the impact of alcohol on cortical development in a dish: strategies from mapping neural stem cell fate. Methods Mol Biol 447:151–168CrossRefPubMedPubMedCentral Miranda RC, Santillano DR, Camarillo C, Dohrman D (2008) Modeling the impact of alcohol on cortical development in a dish: strategies from mapping neural stem cell fate. Methods Mol Biol 447:151–168CrossRefPubMedPubMedCentral
go back to reference Morgan DL, Price HC, Fernando R, Chanda SM, O’Connor RW, Barone SS, Herr DW, Beliles RP (2006) Gestational mercury vapour exposure and diet contribute to mercury accumulation in neonatal rats. Environ Health Perspect 114:735–739CrossRefPubMedPubMedCentral Morgan DL, Price HC, Fernando R, Chanda SM, O’Connor RW, Barone SS, Herr DW, Beliles RP (2006) Gestational mercury vapour exposure and diet contribute to mercury accumulation in neonatal rats. Environ Health Perspect 114:735–739CrossRefPubMedPubMedCentral
go back to reference Patnaik R, Padhy RN (2015) Cellular and nuclear toxicity of HgCl2 to in vitro grown lymphocytes from human umbilical cord blood. Proc Natl Acad Sci India Sec B Biol Sci 85:821–830CrossRef Patnaik R, Padhy RN (2015) Cellular and nuclear toxicity of HgCl2 to in vitro grown lymphocytes from human umbilical cord blood. Proc Natl Acad Sci India Sec B Biol Sci 85:821–830CrossRef
go back to reference Patnaik R, Padhy RN (2016) Evaluation of geno-toxicity of methyl parathion and chlorpyrifos to human liver carcinoma cell line (HepG2). Environ Sci Poll Res 23:8492–8499CrossRef Patnaik R, Padhy RN (2016) Evaluation of geno-toxicity of methyl parathion and chlorpyrifos to human liver carcinoma cell line (HepG2). Environ Sci Poll Res 23:8492–8499CrossRef
go back to reference Petroni D, Tsai J, Mondal D, George W (2013) Attenuation of low dose methylmercury and glutamate induced-cytotoxicity and tau phosphorylation by an N-methyl-D-aspartate antagonist in human neuroblastoma (SHSY5Y) cells. Environ Toxicol 28:700–706CrossRefPubMed Petroni D, Tsai J, Mondal D, George W (2013) Attenuation of low dose methylmercury and glutamate induced-cytotoxicity and tau phosphorylation by an N-methyl-D-aspartate antagonist in human neuroblastoma (SHSY5Y) cells. Environ Toxicol 28:700–706CrossRefPubMed
go back to reference Pluchino S, Quattrini A, Brambilla E, Gritti A, Salani G, Dina G, Galli R, Del Carro U, Amadio S, Bergami A, Furlan R, Comi G, Vescovi AL, Martino G (2003) Injection of adult neurospheres induces recovery in a chronic model of multiple sclerosis. Nature 422:688–694CrossRefPubMed Pluchino S, Quattrini A, Brambilla E, Gritti A, Salani G, Dina G, Galli R, Del Carro U, Amadio S, Bergami A, Furlan R, Comi G, Vescovi AL, Martino G (2003) Injection of adult neurospheres induces recovery in a chronic model of multiple sclerosis. Nature 422:688–694CrossRefPubMed
go back to reference Pistollato F, Bremer-Hoffmann S, Healy L, Young L, Stacey G (2012) Standardization of pluripotent stem cell cultures for toxicity testing. Expert Opin Drug Metab Toxicol 8:239–257CrossRefPubMed Pistollato F, Bremer-Hoffmann S, Healy L, Young L, Stacey G (2012) Standardization of pluripotent stem cell cultures for toxicity testing. Expert Opin Drug Metab Toxicol 8:239–257CrossRefPubMed
go back to reference Rath S, Sahu MC, Dubey D, Debata NK, Padhy RN (2011) Which value should be used as the lethal concentration 50 (LC50) with bacteria? Interdisc Sci: Comput Life Sci 3:138–143CrossRef Rath S, Sahu MC, Dubey D, Debata NK, Padhy RN (2011) Which value should be used as the lethal concentration 50 (LC50) with bacteria? Interdisc Sci: Comput Life Sci 3:138–143CrossRef
go back to reference Smirnova HT, Hogberg M, Leist Hartung T (2014) Developmental neurotoxicity-challenges in the 21st century and in vitro opportunities. ALTEX 31:129–156PubMedPubMedCentral Smirnova HT, Hogberg M, Leist Hartung T (2014) Developmental neurotoxicity-challenges in the 21st century and in vitro opportunities. ALTEX 31:129–156PubMedPubMedCentral
go back to reference Scott CW, Peters MF, Dragan YP (2013) Human induced pluripotent stem cells and their use in drug discovery for toxicity testing. Toxicol Lett 219:49–58CrossRefPubMed Scott CW, Peters MF, Dragan YP (2013) Human induced pluripotent stem cells and their use in drug discovery for toxicity testing. Toxicol Lett 219:49–58CrossRefPubMed
go back to reference Storch A, Schwarz J (2002) Neural stem cells and neurodegeneration. Curr Opin Investig Drug 3:774–781 Storch A, Schwarz J (2002) Neural stem cells and neurodegeneration. Curr Opin Investig Drug 3:774–781
go back to reference Sun W, Buzanska L, Salvi R, Domanska-Janik K, Stachowiak MK (2005) Voltage- sensitive and ligand-gated channels in differentiating fetal neural stem cells isolated from the non-hematopoietic fraction of human umbilical cord blood. Stem Cell 23:93–102CrossRef Sun W, Buzanska L, Salvi R, Domanska-Janik K, Stachowiak MK (2005) Voltage- sensitive and ligand-gated channels in differentiating fetal neural stem cells isolated from the non-hematopoietic fraction of human umbilical cord blood. Stem Cell 23:93–102CrossRef
go back to reference Tchounwou PB, Ayensu WK, Ninashvili N, Sutton D (2003) Environmental exposure to mercury and its toxicopathologic implications for public health. Environ Toxicol 18:149–175CrossRefPubMed Tchounwou PB, Ayensu WK, Ninashvili N, Sutton D (2003) Environmental exposure to mercury and its toxicopathologic implications for public health. Environ Toxicol 18:149–175CrossRefPubMed
go back to reference Zahir F, Rizwi SJ, Haqb SK, Khan RH (2005) Low dose mercury toxicity and human health. Environ Toxicol Pharmacol 20:351–360CrossRefPubMed Zahir F, Rizwi SJ, Haqb SK, Khan RH (2005) Low dose mercury toxicity and human health. Environ Toxicol Pharmacol 20:351–360CrossRefPubMed
go back to reference Zigova T, Song S, Willing AE, Hudson JE, Newman MB, Saporta S, Sanchez-Ramos J, Sanberg PR (2002) Human umbilical cord blood cells express neural antigens after transplantation into the developing rat brain. Cell Transplant 11:265–274CrossRefPubMed Zigova T, Song S, Willing AE, Hudson JE, Newman MB, Saporta S, Sanchez-Ramos J, Sanberg PR (2002) Human umbilical cord blood cells express neural antigens after transplantation into the developing rat brain. Cell Transplant 11:265–274CrossRefPubMed
Metadata
Title
Human Umbilical Cord Blood-Derived Neural Stem Cell Line as a Screening Model for Toxicity
Authors
Rajashree Patnaik
Rabindra Nath Padhy
Publication date
01-04-2017
Publisher
Springer US
Published in
Neurotoxicity Research / Issue 3/2017
Print ISSN: 1029-8428
Electronic ISSN: 1476-3524
DOI
https://doi.org/10.1007/s12640-016-9681-x

Other articles of this Issue 3/2017

Neurotoxicity Research 3/2017 Go to the issue