Skip to main content

Advertisement

Springer Nature Link
Log in

Effects of mono-(2-ethylhexyl) phthalate (MEHP) on chicken germ cells cultured in vitro

  • Ecotoxicology and Environmental Toxicology : new concepts, new tools
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

In recent decades, many toxicological tests based on in vivo or in vitro models, mainly from mammalian (rat–mouse) and fish species, were used to assess the risks raised by contact or ingestion of molecules of pharmaceutical, agricultural, or natural origin. But no, or few, in vitro tests using other non-mammalian models such as bird have been explored despite their advantages: the embryonic gonads of birds have a high plasticity of development sensitive to estrogen, and sperm production is nearly two times faster than in rodents. Hence, we have established an in vitro culture of germ cells and somatic cells from chicken post-natal testis, and we have evaluated the sensitivity against the endocrine disruptor compound mono-(2-ethylhexyl) phthalate (MEHP) in comparison to previous studies using rodent and human models. After 96 h of exposure in presence of 10 μM MEHP, chicken seminiferous tubules cultures present a structural alteration, a reduction in cell proliferation and in germ cells population. Apoptosis of germ and somatic cells increases in presence of 1 μM MEHP. Furthermore, MEHP does not affect inhibin B and lactate production by Sertoli cells. These results are in accordance with previous studies using rat, mice, or human culture of testicular cells and in similar range of exposures or even better sensitivity for some “end-points” (biological parameters). In conclusion, the establishment of this postnatal testicular cells culture could be considered as an alternative method to in vivo experiments frequently used for evaluating the impact on the terrestrial wildlife species. This method could be also complementary to mammal model due to the limiting number of animals used and its elevated sensitivity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Alam MS, Ohsako S, Tay TW, Tsunekawa N, Kanai Y, Kurohmaru M (2010) Di(n-butyl) phthalate induces vimentin filaments disruption in rat sertoli cells: a possible relation with spermatogenic cell apoptosis. Anat Histol Embryol 39:186–193

    Article  CAS  Google Scholar 

  • Auharek SA, de Franca LR, McKinnell C, Jobling MS, Scott HM, Sharpe RM (2010) Prenatal plus postnatal exposure to di(n-butyl) phthalate and/or flutamide markedly reduces final Sertoli cell number in the rat. Endocrinology 151:2868–2875

    Article  CAS  Google Scholar 

  • Awal MA, Kurohmaru M, Ishii M, Andriana BB, Kanai Y, Hayashi Y (2004) Mono-(2-ethylhexyl) phthalate (MEHP) induces spermatogenic cell apoptosis in guinea pig testes at prepubertal stage in vitro. Int J Toxicol 23:349–355

    Article  CAS  Google Scholar 

  • Bakst MR, Akuffo V, Trefil P, Brillard JP (2007) Morphological and histochemical characterization of the seminiferous epithelial and Leydig cells of the turkey. Anim Reprod Sci 97:303–313

    Article  CAS  Google Scholar 

  • Blomqvist A, Berg C, Holm L, Brandt I, Ridderstrale Y, Brunstrom B (2006) Defective reproductive organ morphology and function in domestic rooster embryonically exposed to o, p′-DDT or ethynylestradiol. Biol Reprod 74:481–486

    Article  CAS  Google Scholar 

  • Blount BC, Milgram KE, Silva MJ, Malek NA, Reidy JA, Needham LL, Brock JW (2000) Quantitative detection of eight phthalate metabolites in human urine using HPLC–APCI–MS/MS. Anal Chem 72:4127–4134

    Article  CAS  Google Scholar 

  • Bobes RJ, Castro JI, Miranda C, Romano MC (2001) Insulin modifies the proliferation and function of chicken testis cells. Poult Sci 80:637–642

    CAS  Google Scholar 

  • Carreau S, Silandre D, Bois C, Bouraima H, Galeraud-Denis I, Delalande C (2007) Estrogens: a new player in spermatogenesis. Folia Histochem Cytobiol 45(Suppl 1):S5–S10

    Google Scholar 

  • Chauvigne F, Menuet A, Lesne L, Chagnon MC, Chevrier C, Regnier JF, Angerer J, Jegou B (2009) Time- and dose-related effects of di-(2-ethylhexyl) phthalate and its main metabolites on the function of the rat fetal testis in vitro. Environ Health Perspect 117:515–521

    CAS  Google Scholar 

  • Creasy DM, Beech LM, Gray TJ (1988) Effects of mono-(2-ethylhexyl) phthalate and mono-n-pentyl phthalate on the ultrastructural morphology of rat Sertoli cells in Sertoli/germ cell co-cultures: correlation with the in vivo effects of di-n-pentyl phthalate. Toxicol In Vitro 2:83–95

    Article  CAS  Google Scholar 

  • Curto KA, Thomas JA (1982) Comparative effects of diethylhexyl phthalate or monoethylhexyl phthalate on male mouse and rat reproductive organs. Toxicol Appl Pharmacol 62:121–125

    Article  CAS  Google Scholar 

  • Dalgaard M, Nellemann C, Lam HR, Sorensen IK, Ladefoged O (2001) The acute effects of mono(2-ethylhexyl)phthalate (MEHP) on testes of prepubertal Wistar rats. Toxicol Lett 122:69–79

    Article  CAS  Google Scholar 

  • de Reviers M, Hochereau-de Reviers MT, Blanc MR, Brillard JP, Courot M, Pelletier J (1980) Control of Sertoli and germ cell populations in the cock and sheep testes. Reprod Nutr Dev 20:241–249

    Article  Google Scholar 

  • Desdoits-Lethimonier C, Albert O, Le Bizec B, Perdu E, Zalko D, Courant F, Lesne L, Guille F, Dejucq-Rainsford N, Jegou B (2012) Human testis steroidogenesis is inhibited by phthalates. Hum Reprod 27:1451–1459

    Article  CAS  Google Scholar 

  • Djakiew D, Dym M (1988) Pachytene spermatocyte proteins influence Sertoli cell function. Biol Reprod 39:1193–1205

    Article  CAS  Google Scholar 

  • Ellingson DJ, Yao KT (1970) Growth and observations of Chinese hamster seminiferous epithelium in vitro. J Cell Sci 6:195–205

    CAS  Google Scholar 

  • Erkekoglu P, Rachidi W, Yuzugullu OG, Giray B, Favier A, Ozturk M, Hincal F (2010) Evaluation of cytotoxicity and oxidative DNA damaging effects of di(2-ethylhexyl)-phthalate (DEHP) and mono(2-ethylhexyl)-phthalate (MEHP) on MA-10 Leydig cells and protection by selenium. Toxicol Appl Pharmacol 248:52–62

    Article  CAS  Google Scholar 

  • Fisher JS, Macpherson S, Marchetti N, Sharpe RM (2003) Human “testicular dysgenesis syndrome”: a possible model using in-utero exposure of the rat to dibutyl phthalate. Hum Reprod 18:1383–1394

    Article  CAS  Google Scholar 

  • Foster PM, Creasy DM, Foster JR, Gray TJ (1984) Testicular toxicity produced by ethylene glycol monomethyl and monoethyl ethers in the rat. Environ Health Perspect 57:207–217

    Article  CAS  Google Scholar 

  • Froment P, Dupont J, Christophe-Marine J (2008) Mdm2 exerts pro-apoptotic activities by antagonizing insulin-like growth factor-I-mediated survival. Cell Cycle 7:3098–3103

    Article  CAS  Google Scholar 

  • Fry DM (1995) Reproductive effects in birds exposed to pesticides and industrial chemicals. Environ Health Perspect 103(Suppl 7):165–171

    Article  CAS  Google Scholar 

  • Gray TJ (1986) Testicular toxicity in vitro: Sertoli–germ cell co-cultures as a model system. Food Chem Toxicol 24:601–605

    Article  CAS  Google Scholar 

  • Gray TJ, Beamand JA (1984) Effect of some phthalate esters and other testicular toxins on primary cultures of testicular cells. Food Chem Toxicol 22:123–131

    Article  CAS  Google Scholar 

  • Gray TJ, Rowland IR, Foster PM, Gangolli SD (1982) Species differences in the testicular toxicity of phthalate esters. Toxicol Lett 11:141–147

    Article  CAS  Google Scholar 

  • Guibert E, Briere S, Pelletier R, Brillard JP, Froment P (2011) Characterization of chicken Sertoli cells in vitro. Poult Sci 90:1276–1286

    Article  CAS  Google Scholar 

  • Guillette LJ Jr, Gross TS, Masson GR, Matter JM, Percival HF, Woodward AR (1994) Developmental abnormalities of the gonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida. Environ Health Perspect 102:680–688

    Article  CAS  Google Scholar 

  • Hauser R, Meeker JD, Singh NP, Silva MJ, Ryan L, Duty S, Calafat AM (2007) DNA damage in human sperm is related to urinary levels of phthalate monoester and oxidative metabolites. Hum Reprod 22:688–695

    Article  CAS  Google Scholar 

  • Hogberg J, Hanberg A, Berglund M, Skerfving S, Remberger M, Calafat AM, Filipsson AF, Jansson B, Johansson N, Appelgren M, Hakansson H (2008) Phthalate diesters and their metabolites in human breast milk, blood or serum, and urine as biomarkers of exposure in vulnerable populations. Environ Health Perspect 116:334–339

    Article  Google Scholar 

  • Huleihel M, Lunenfeld E (2004) Regulation of spermatogenesis by paracrine/autocrine testicular factors. Asian J Androl 6:259–268

    CAS  Google Scholar 

  • Ichimura T, Kawamura M, Mitani A (2003) Co-localized expression of FasL, Fas, Caspase-3 and apoptotic DNA fragmentation in mouse testis after oral exposure to di(2-ethylhexyl)phthalate. Toxicology 194:35–42

    Article  CAS  Google Scholar 

  • Kijima K, Toyosawa K, Yasuba M, Matsuoka N, Adachi T, Komiyama M, Mori C (2004) Gene expression analysis of the rat testis after treatment with di(2-ethylhexyl) phthalate using cDNA microarray and real-time RT-PCR. Toxicol Appl Pharmacol 200:103–110

    Article  CAS  Google Scholar 

  • Kleymenova E, Swanson C, Boekelheide K, Gaido KW (2005) Exposure in utero to di(n-butyl) phthalate alters the vimentin cytoskeleton of fetal rat Sertoli cells and disrupts Sertoli cell–gonocyte contact. Biol Reprod 73:482–490

    Article  CAS  Google Scholar 

  • Koch HM, Rossbach B, Drexler H, Angerer J (2003) Internal exposure of the general population to DEHP and other phthalates—determination of secondary and primary phthalate monoester metabolites in urine. Environ Res 93:177–185

    Article  CAS  Google Scholar 

  • Kwon S, Hess RA, Bunick D, Nitta H, Janulis L, Osawa Y, Bahr JM (1995) Rooster testicular germ cells and epididymal sperm contain P450 aromatase. Biol Reprod 53:1259–1264

    Article  CAS  Google Scholar 

  • Lambrot R, Muczynski V, Lecureuil C, Angenard G, Coffigny H, Pairault C, Moison D, Frydman R, Habert R, Rouiller-Fabre V (2009) Phthalates impair germ cell development in the human fetal testis in vitro without change in testosterone production. Environ Health Perspect 117:32–37

    Article  CAS  Google Scholar 

  • Latini G, Del Vecchio A, Massaro M, Verrotti A, De Felice C (2006) Phthalate exposure and male infertility. Toxicology 226:90–98

    Article  CAS  Google Scholar 

  • Lavial F, Acloque H, Bachelard E, Nieto MA, Samarut J, Pain B (2009) Ectopic expression of Cvh (Chicken Vasa homologue) mediates the reprogramming of chicken embryonic stem cells to a germ cell fate. Dev Biol 330:73–82

    Article  CAS  Google Scholar 

  • Lee SK, Veeramachaneni DN (2005) Subchronic exposure to low concentrations of di-n-butyl phthalate disrupts spermatogenesis in Xenopus laevis frogs. Toxicol Sci 84:394–407

    Article  CAS  Google Scholar 

  • Lee YM, Jung JG, Kim JN, Park TS, Kim TM, Shin SS, Kang DK, Lim JM, Han JY (2006) A testis-mediated germline chimera production based on transfer of chicken testicular cells directly into heterologous testes. Biol Reprod 75:380–386

    Article  CAS  Google Scholar 

  • Legendre A, Froment P, Desmots S, Lecomte A, Habert R, Lemazurier E (2010) An engineered 3D blood–testis barrier model for the assessment of reproductive toxicity potential. Biomaterials 31:4492–4505

    Article  CAS  Google Scholar 

  • Lehraiki A, Racine C, Krust A, Habert R, Levacher C (2009) Phthalates impair germ cell number in the mouse fetal testis by an androgen- and estrogen-independent mechanism. Toxicol Sci 111:372–382

    Article  CAS  Google Scholar 

  • Li H, Kim KH (2003) Effects of mono-(2-ethylhexyl) phthalate on fetal and neonatal rat testis organ cultures. Biol Reprod 69:1964–1972

    Article  CAS  Google Scholar 

  • Li B, Wang X-Y, Tian Z, Xiao X-J, Xu Q, Wei C-X, Y F, Sun H-C, Chen G-H (2010) Directional differentiation of chicken spermatogonial stem cells in vitro. Cytotherapy 12:326–331

    Google Scholar 

  • Lucas BE, Fields C, Joshi N, Hofmann MC (2012) Mono-(2-ethylhexyl)-phthalate (MEHP) affects ERK-dependent GDNF signalling in mouse stem-progenitor spermatogonia. Toxicology 299:10–19

    Article  CAS  Google Scholar 

  • Mi Y, Zhang C, Zeng W, Tang X (2005) Establishment of a germ-somatic cell coculture model for toxicity assessment of environmental endocrine disrupters. J Environ Sci Health, Part A: Tox Hazard Subst Environ Eng 40:1917–1928

    Article  CAS  Google Scholar 

  • Mi YL, Zhang CQ, Zeng WD, Liu JX, Liu HY (2007) The isoflavonoid daidzein attenuates the oxidative damage induced by polychlorinated biphenyls on cultured chicken testicular cells. Poult Sci 86:2008–2012

    CAS  Google Scholar 

  • Migliorini D, Lazzerini Denchi E, Danovi D, Jochemsen A, Capillo M, Gobbi A, Helin K, Pelicci PG, Marine JC (2002) Mdm4 (Mdmx) regulates p53-induced growth arrest and neuronal cell death during early embryonic mouse development. Mol Cell Biol 22:5527–5538

    Article  CAS  Google Scholar 

  • Moss EJ, Cook MW, Thomas LV, Gray TJ (1988) The effect of mono-(2-ethylhexyl) phthalate and other phthalate esters on lactate production by Sertoli cells in vitro. Toxicol Lett 40:77–84

    Article  CAS  Google Scholar 

  • Muczynski V, Cravedi JP, Lehraiki A, Levacher C, Moison D, Lecureuil C, Messiaen S, Perdu E, Frydman R, Habert R, Rouiller-Fabre V (2012) Effect of mono-(2-ethylhexyl) phthalate on human and mouse fetal testis: in vitro and in vivo approaches. Toxicol Appl Pharmacol 261:97–104

    Article  CAS  Google Scholar 

  • Mylchreest E, Sar M, Wallace DG, Foster PM (2002) Fetal testosterone insufficiency and abnormal proliferation of Leydig cells and gonocytes in rats exposed to di(n-butyl) phthalate. Reprod Toxicol 16:19–28

    Article  CAS  Google Scholar 

  • O’Donnell L, Narula A, Balourdos G, Gu YQ, Wreford NG, Robertson DM, Bremner WJ, McLachlan RI (2001) Impairment of spermatogonial development and spermiation after testosterone-induced gonadotropin suppression in adult monkeys (Macaca fascicularis). J Clin Endocrinol Metab 86:1814–1822

    Article  Google Scholar 

  • Opalka M, Kaminska B, Ciereszko R, Dusza L (2004) Genistein affects testosterone secretion by Leydig cells in roosters (Gallus gallus domesticus). Reprod Biol 4:185–193

    Google Scholar 

  • Orth JM, Gunsalus GL, Lamperti AA (1988) Evidence from Sertoli cell-depleted rats indicates that spermatid number in adults depends on numbers of Sertoli cells produced during perinatal development. Endocrinology 122:787–794

    Article  CAS  Google Scholar 

  • Rasoulpour RJ, Boekelheide K (2005) NF-kappaB is activated in the rat testis following exposure to mono-(2-ethylhexyl) phthalate. Biol Reprod 72:479–486

    Article  CAS  Google Scholar 

  • Rattner BA, Eroschenko VP, Fox GA, Fry DM, Gorsline J (1984) Avian endocrine responses to environmental pollutants. J Exp Zool 232:683–689

    Article  CAS  Google Scholar 

  • Richburg JH, Boekelheide K (1996) Mono-(2-ethylhexyl) phthalate rapidly alters both Sertoli cell vimentin filaments and germ cell apoptosis in young rat testes. Toxicol Appl Pharmacol 137:42–50

    Article  CAS  Google Scholar 

  • Richburg JH, Nanez A, Williams LR, Embree ME, Boekelheide K (2000) Sensitivity of testicular germ cells to toxicant-induced apoptosis in gld mice that express a nonfunctional form of Fas ligand. Endocrinology 141:787–793

    Article  CAS  Google Scholar 

  • Rogers R, Ouellet G, Brown C, Moyer B, Rasoulpour T, Hixon M (2008) Cross-talk between the Akt and NF-kappaB signaling pathways inhibits MEHP-induced germ cell apoptosis. Toxicol Sci 106:497–508

    Article  CAS  Google Scholar 

  • Rosado-Berrios CA, Velez C, Zayas B (2011) Mitochondrial permeability and toxicity of diethylhexyl and monoethylhexyl phthalates on TK6 human lymphoblasts cells. Toxicol In Vitro 25:2010–2016

    Article  CAS  Google Scholar 

  • Rosenstrauch A, Degen AA, Friedlander M (1994) Spermatozoa retention by Sertoli cells during the decline in fertility in aging roosters. Biol Reprod 50:129–136

    Article  CAS  Google Scholar 

  • Scott HM, Hutchison GR, Mahood IK, Hallmark N, Welsh M, De Gendt K, Verhoeven G, O’Shaughnessy P, Sharpe RM (2007) Role of androgens in fetal testis development and dysgenesis. Endocrinology 148:2027–2036

    Article  CAS  Google Scholar 

  • Sekercioglu CH, Daily GC, Ehrlich PR (2004) Ecosystem consequences of bird declines. Proc Natl Acad Sci U S A 101:18042–18047

    Article  CAS  Google Scholar 

  • Sheweita SA, Tilmisany AM, Al-Sawaf H (2005) Mechanisms of male infertility: role of antioxidants. Curr Drug Metab 6:495–501

    Article  CAS  Google Scholar 

  • Skakkebaek NE, Jorgensen N, Main KM, Rajpert-De Meyts E, Leffers H, Andersson AM, Juul A, Carlsen E, Mortensen GK, Jensen TK, Toppari J (2006) Is human fecundity declining? Int J Androl 29:2–11

    Article  Google Scholar 

  • Smits JE, Fernie KJ (2012) Avian wildlife as sentinels of ecosystem health. Comp Immunol Microbiol Infect Dis. doi:10.1016/j.cimid.2012.11.007

  • Sobarzo CM, Lustig L, Ponzio R, Suescun MO, Denduchis B (2009) Effects of di(2-ethylhexyl) phthalate on gap and tight junction protein expression in the testis of prepubertal rats. Microsc Res Tech 72:868–877

    Article  CAS  Google Scholar 

  • Staub C, Hue D, Nicolle JC, Perrard-Sapori MH, Segretain D, Durand P (2000) The whole meiotic process can occur in vitro in untransformed rat spermatogenic cells. Exp Cell Res 260:85–95

    Article  CAS  Google Scholar 

  • Swan SH, Main KM, Liu F, Stewart SL, Kruse RL, Calafat AM, Mao CS, Redmon JB, Ternand CL, Sullivan S, Teague JL (2005) Decrease in anogenital distance among male infants with prenatal phthalate exposure. Environ Health Perspect 113:1056–1061

    Article  CAS  Google Scholar 

  • Toebosch AM, Brussee R, Verkerk A, Grootegoed JA (1989) Quantitative evaluation of the maintenance and development of spermatocytes and round spermatids in cultured tubule fragments from immature rat testis. Int J Androl 12:360–374

    Article  CAS  Google Scholar 

  • Tsunekawa N, Naito M, Sakai Y, Nishida T, Noce T (2000) Isolation of chicken vasa homolog gene and tracing the origin of primordial germ cells. Development 127:2741–2750

    CAS  Google Scholar 

  • Uren-Webster TM, Lewis C, Filby AL, Paull GC, Santos EM (2010) Mechanisms of toxicity of di(2-ethylhexyl) phthalate on the reproductive health of male zebrafish. Aquat Toxicol 99:360–369

    Article  CAS  Google Scholar 

  • Vos JG, Dybing E, Greim HA, Ladefoged O, Lambre C, Tarazona JV, Brandt I, Vethaak AD (2000) Health effects of endocrine-disrupting chemicals on wildlife, with special reference to the European situation. Crit Rev Toxicol 30:71–133

    Article  CAS  Google Scholar 

  • Williams J, Foster PM (1988) The production of lactate and pyruvate as sensitive indices of altered rat Sertoli cell function in vitro following the addition of various testicular toxicants. Toxicol Appl Pharmacol 94:160–170

    Article  CAS  Google Scholar 

  • Yao PL, Lin YC, Richburg JH (2010) Mono-(2-ethylhexyl) phthalate-induced disruption of junctional complexes in the seminiferous epithelium of the rodent testis is mediated by MMP2. Biol Reprod 82:516–527

    Article  CAS  Google Scholar 

  • Zhang YH, Lin L, Liu ZW, Jiang XZ, Chen BH (2008) Disruption effects of monophthalate exposures on inter-Sertoli tight junction in a two-compartment culture model. Environ Toxicol 23:302–308

    Article  CAS  Google Scholar 

  • Zhu XB, Tay TW, Andriana BB, Alam MS, Choi EK, Tsunekawa N, Kanai Y, Kurohmaru M (2010) Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice. Okajimas Folia Anat Jpn 86:129–136

    Article  CAS  Google Scholar 

  • Zwain IH, Cheng CY (1994) Rat seminiferous tubular culture medium contains a biological factor that inhibits Leydig cell steroidogenesis: its purification and mechanism of action. Mol Cell Endocrinol 104:213–227

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Institut National de la Recherche Agronomique, INRA, and by the French Ministry of Environment in a national program Grenelle de l’Environnement programme LOLF 189 funding (Specific Tools and Methods for Reprotoxicity program).

We thank Frederic Mercerand, Jean Marie Brigant, Harold Rigoreau, and the team for expert animal care.

We are grateful to Daniele Carré-Eusèbe, Emmanuelle Oréal, and Jean-Yves Picard (Paris, France) for generously providing us the antibody against chAMH.

We thank Xavier Druart and Guillaume Tsikis for their expertise on the flow cytometer.

Conflict of interest

The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work.

Author disclosure summary

DISCLOSURE STATEMENT: The authors have nothing to disclose.

Author information

Authors and Affiliations

  1. UMR 6175 INRA CNRS Université de Tours Haras Nationaux Physiologie de la Reproduction et des Comportements, 37380, Nouzilly, France

    Edith Guibert, Bérénice Prieur & Pascal Froment

  2. LUNAM Université, ONIRIS, USC 1329 INRA Laboratoire d’Etude des Résidus et Contaminants dans les Aliments (LABERCA), F-44307, Nantes, France

    Ronan Cariou, Frédérique Courant & Jean Philippe Antignac

  3. INSERM, U846, Stem Cell and Brain Research Institute, Bron, France

    Bertrand Pain

  4. INRA, USC1361, AGROBIOSTEM, Bron, France

    Bertrand Pain

  5. Université de Lyon, Lyon 1, UMR S 846, Lyon, France

    Bertrand Pain

  6. FERTIL’AVI, 10 rue du 8 Mai, 37360, Rouziers de Touraine, France

    Jean Pierre Brillard

  7. UMR 6175 INRA CNRS Université de Tours Haras Nationaux Physiologie de la Reproduction et des Comportements, Station de Recherche Avicole, 37380, Nouzilly, France

    Pascal Froment

Authors
  1. Edith Guibert
    View author publications

    Search author on:PubMed Google Scholar

  2. Bérénice Prieur
    View author publications

    Search author on:PubMed Google Scholar

  3. Ronan Cariou
    View author publications

    Search author on:PubMed Google Scholar

  4. Frédérique Courant
    View author publications

    Search author on:PubMed Google Scholar

  5. Jean Philippe Antignac
    View author publications

    Search author on:PubMed Google Scholar

  6. Bertrand Pain
    View author publications

    Search author on:PubMed Google Scholar

  7. Jean Pierre Brillard
    View author publications

    Search author on:PubMed Google Scholar

  8. Pascal Froment
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Pascal Froment.

Additional information

Responsible editor: Philippe Garrigues

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guibert, E., Prieur, B., Cariou, R. et al. Effects of mono-(2-ethylhexyl) phthalate (MEHP) on chicken germ cells cultured in vitro. Environ Sci Pollut Res 20, 2771–2783 (2013). https://doi.org/10.1007/s11356-013-1487-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-013-1487-2

Keywords

Profiles

  1. Bertrand Pain View author profile