Skip to main content
SpringerLink
Log in

Leptin treatment of in vitro cultured embryos increases outgrowth rate of inner cell mass during embryonic stem cell derivation

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Leptin, a metabolic hormone, regulates the reproductive functions responding to both nutritional and body conditions. Embryonic stem cells play important roles in reproductive technology, but their derivation can be challenging. In this study, we evaluated the derivation rates of mouse embryonic stem cell (mESC) line from blastocysts developing in embryo culture media supplemented with different leptin concentrations. The results showed that addition of leptin into the embryo culture medium supported the in vitro development of mouse embryo. The mESC line derivation rates for media treated with 0, 10, 50, and 100 ng/ml of leptin were 61.24 % (54/88), 84.96 % (42/50), 81.79 % (61/76), and 85.78 % (56/67), respectively. In addition, leptin treatment of blastocysts upregulated the expression levels of the trophectoderm marker Cdx2, whereas inner cell mass markers Oct-4 and Nanog were not affected. mESC lines derived after leptin treatment demonstrated hallmarks of pluripotency, such as alkaline phosphatase activity, expression of, OCT4, NANOG, and SSEA1, as well as the ability to form embryoid bodies and well-differentiated teratomas. In conclusion, leptin has a positive effect on the derivation rate of mouse embryonic stem cell lines which may be, in part, due to its effects on the development of the trophectoderm cell lineage in the embryo.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3

Similar content being viewed by others

References

  • Bagis H, Akkoc T, Taskin C, Arat S (2010) Comparison of different cryopreservation techniques: higher survival and implantation rate of frozen–thawed mouse pronuclear embryos in the presence of beta-mercaptoethanol in post-thaw culture. Reprod Domest Anim 45:332–337

    Article  CAS  Google Scholar 

  • Cobler L, Pera M, Garrido M, Iglesias M, de Bolós C (2014) CDX2 can be regulated through the signalling pathways activated by IL-6 in gastric cells. Biochim Biophys Acta 1839:785–792

    Article  CAS  PubMed  Google Scholar 

  • Coskun M, Olsen AK, Bzorek M, Holck S, Engel UH, Nielsen OH, Troelsen JT (2014) Involvement of CDX2 in the cross talk between TNF-α and Wnt signaling pathway in the colon cancer cell line Caco-2. Carcinogenesis 35:1185–1192

    Article  CAS  PubMed  Google Scholar 

  • Costa-Borges N, Santaló J, Ibáñez E (2010) Comparison between the effect of valproic acid and trichostatin A on the in vitro development, blastocyst quality, and full-term development of mouse somatic cell nuclear transfer embryos. Cell Reprogram 12(4):437–46

  • Czechanski A, Byers C, Greenstein I, Schrode N, Donahue LR, Hadjantonakis AK, Reinholdt L (2014) Derivation and characterization of mouse embryonic stem cells from permissive and nonpermissive strains. Nat Protoc 9:559–574

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Evans MJ, Kaufman MF (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156

    Article  CAS  Google Scholar 

  • Fedorcsák P, Dale PO, Storeng R, Ertzeid G, Bjercke S, Oldereid N, Omland AK, Åbyholm T, Tanbo T (2004) Impact of overweight and underweight on assisted reproduction treatment. Hum Reprod 19:2523–2528

    Article  PubMed  Google Scholar 

  • Feldman DE, Chen C, Punj V, Tsukamoto H, Machida K (2012) Pluripotency factor-mediated expression of the leptin receptor (OB-R) links obesity to oncogenesis through tumor-initiating stem cells. Proc Natl Acad Sci U S A 109:829–834

    Article  PubMed  Google Scholar 

  • Fidan K, Kavaklıoğlu G, Ebrahimi A, Özlü C, Ay NZ, Ruacan A, Gül A, Önder TT (2015) Generation of integration-free induced pluripotent stem cells from a patient with Familial Mediterranean Fever (FMF). Stem Cell Res 15:694–696

    Article  CAS  PubMed  Google Scholar 

  • Herrid M, Nguyen VL, Hinch G, McFarlane JR (2006) Leptin has concentration and stage-dependent effects on embryonic development in vitro. Reproduction 132:247–256

    Article  CAS  PubMed  Google Scholar 

  • Iijima S, Tanimoto Y, Mizuno S, Daitoku Y, Kunita S, Sugiyama F, Yagami KI (2010) Effect of different culture conditions on establishment of embryonic stem cells from BALB/cAJ and NZB/BINJ mice. Cell Rep 12:679–688

    Article  CAS  Google Scholar 

  • Inagaki-Ohara K, Okamoto S, Takagi K, Saito K, Arita S, Tang L, Hori T, Kataoka H, Matsumoto S, Minokoshi Y (2016) Leptin receptor signaling is required for high-fat diet-induced atrophic gastritis in mice. Nutr Metab 13:7

    Article  CAS  Google Scholar 

  • Kanda A, Sotomaru Y, Shiozawa S, Hiyama E (2012) Establishment of ES cells from inbred strain mice by dual inhibition (2i). J Reprod Dev 58:77–83

    Article  CAS  PubMed  Google Scholar 

  • Kawamura K, Sato N, Fukuda J, Kodama H, Kumagai J, Tanikawa H, Murata M, Tanaka T (2003) The role of leptin during the development of mouse preimplantation embryos. Mol Cell Endocrinol 202:185–189

    Article  CAS  PubMed  Google Scholar 

  • Kawamura K, Sato N, Fukuda J, Kodama H, Kumagai J, Tanikawa H, Nakamura A, Tanaka T (2002) Leptin promotes the development of mouse preimplantation embryos in vitro. Endocrinology 143:1922–1931

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Schoonjans L, Tielens S, Speleman F, Cornelissen M, De Sutter P, Dhont MARC, Van der Elst J (2006) Culturing in vitro produced blastocysts in sequential media promotes ES cell derivation. Mol Reprod Dev 73:1017–1021

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • Lu CW, Yabuuchi A, Chen L, Viswanathan S, Kim K, Daley GQ (2008) Ras-MAPK signaling promotes trophectoderm formation from embryonic stem cells and mouse embryos. Nat Genet 40(7):921–926

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma Z, Swigut T, Valouev A, Rada-Iglesias A, Wysocka J (2011) Sequence-specific regulator Prdm14 safeguards mouse ESCs from entering extraembryonic endoderm fates. Nat Struct Mol Biol 18:120–127

    Article  CAS  PubMed  Google Scholar 

  • Mallol A, Santaló J, Ibáñez E (2013) Comparison of three differential mouse blastocyst staining methods. Syst Biol Reprod Med 59:117–122

    Article  CAS  PubMed  Google Scholar 

  • Martin GR (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci US A 78:7634–7638

    Article  CAS  Google Scholar 

  • Nichols J, Chambers I, Smith A (1994) Derivation of germline competent embryonic stem cells with a combination of interleukin-6 and soluble interleukin-6 receptor. Exp Cell Res 215:237–239

    Article  CAS  PubMed  Google Scholar 

  • Nichols J, Evans EP, Smith AG (1990) Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity. Development 110:1341–1348

    CAS  PubMed  Google Scholar 

  • Park HK, Ahima RS (2014) Leptin signaling. F1000 Prime Rep 6:73

    Google Scholar 

  • Robertson EJ (1987) Embryo-derived stem cell lines. Teratocarcinomas and embryonic stem cells a practical approach 71–112

  • Rungsiwiwut R, Rungarunlert S, Numchaisrika P, Pruksananonda K, Techakumphu M, Virutamasen P (2008) Effect of leukemia inhibitory factor (LIF) on the quality of in vitro produced mouse blastocysts and subsequent derivation of embryonic stem (ES) cells. J Med Assoc Thai 91:608–614

    PubMed  Google Scholar 

  • Smith AG (2001) Embryo-derived stem cells: of mice and men. Annu Rev Cell Dev Biol 17:435–462

    Article  CAS  PubMed  Google Scholar 

  • Taskin AC, Akkoc T, Sagirkaya H, Bagis H, Arat S (2016) Comparison of the development of mouse embryos manipulated with different biopsy techniques. Turk J Vet Anim Sci 40:157–162

    Article  Google Scholar 

  • Thouas GA, Korfiatis NA, French AJ, Jones GM, Trounson AO (2001) Simplified technique for differential staining of inner cell mass and trophectoderm cells of mouse and bovine blastocysts. Reprod BioMed Online 3:25–29

    Article  PubMed  Google Scholar 

  • Tielens S, Verhasselt B, Liu J, Dhont MARC, Van der Elst J, Cornelissen M (2006) Generation of embryonic stem cell lines from mouse blastocysts developed in vivo and in vitro: relation to Oct-4 expression. Reproduction 132:59–66

    Article  CAS  PubMed  Google Scholar 

  • Wu G, Gentile L, Do JT, Cantz T, Sutter J, Psathaki K, Araúzo-Bravo MJ, Ortmeier C, Schöler HR (2010a) Efficient derivation of pluripotent stem cells from siRNA-mediated Cdx2-deficient mouse embryos. Stem Cells Dev 20(3):485–493

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu G, Gentile L, Fuchikami T, Sutter J, Psathaki K, Esteves TS, Araúzo-Bravo MJ, Ortmeier C, Verberk G, Abe K, Schöler HR (2010b) Initiation of trophectoderm lineage specification in mouse embryos is independent of Cdx2. Development 134:4159–4169

    Article  CAS  Google Scholar 

  • Yang YJ, Cao YJ, Bo SM, Peng S, Liu WM, Duan EK (2006) Leptin-directed embryo implantation: leptin regulates adhesion and outgrowth of mouse blastocysts and receptivity of endometrial epithelial cells. Anim Reprod Sci 92(1–2):155–167

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM (1994) Positional cloning of the mouse obese gene and its human homologue. Nature 372:425–432

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge use of the services and facilities of the Koç University Research Center for Translational Medicine (KUTTAM), funded by the Republic of Turkey Ministry of Development. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Ministry of Development.

Funding

This research supported by a grant from TUBITAK - The Scientific and Technological Research Council of Turkey (Grant Number: TOVAG 113O223).

Author information

Authors and Affiliations

  1. Embryo Manipulation Laboratory, Animal Research Facility, Research Center For Translational Medicine, Koç University, Rumelifeneri yolu, 34450, Sariyer, Istanbul, Turkey

    Ali Cihan Taskin & Ahmet Kocabay

  2. Molecular Biology and Genetics Department, Faculty of Arts and Sciences, Haliç University, Haliç University, Sutluce Mah., Imrahor Cad., No:82, 34445, Istanbul, Turkey

    Ayyub Ebrahimi

  3. Department of Histology and Embryology, School of Medicine, Koç University, Rumelifeneri yolu, 34450, Sariyer, Istanbul, Turkey

    Sercin Karahuseyinoglu & Gizem Nur Sahin

  4. Department of Moleculer Biology and Genetics, School of Medicine, Koç University, Rumelifeneri Yolu, 34450, Sariyer, Istanbul, Turkey

    Burcu Ozcimen & Tamer T. Onder

  5. Department of Pathology, School of Medicine, Koç University, Rumelifeneri Yolu, 34450, Sariyer, Istanbul, Turkey

    Arzu Ruacan

Authors
  1. Ali Cihan Taskin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ahmet Kocabay
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ayyub Ebrahimi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sercin Karahuseyinoglu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gizem Nur Sahin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Burcu Ozcimen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Arzu Ruacan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tamer T. Onder
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ali Cihan Taskin.

Ethics declarations

All mouse experiments and animal care protocols were approved by the Local Ethics Committee for Animal Experiments (Approval No: 2013-06), Koç University.

Conflict of interest

The authors declare that they have no competing interests.

Additional information

Editor: Tetsuji Okamoto

Prior conference presentation of the submitted: Taskin AC, A Kocabay, TT Onder and A Ebrahimi. (2016) Effect of leptin on derivation rate of mouse embryonic stem (ES) cell line. In TRANSGENIC RESEARCH M 25: 265–265 (13th Transgenic Technology Meeting in Prague, Czech Republic, from March 20th–23th, 2016)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Taskin, A.C., Kocabay, A., Ebrahimi, A. et al. Leptin treatment of in vitro cultured embryos increases outgrowth rate of inner cell mass during embryonic stem cell derivation. In Vitro Cell.Dev.Biol.-Animal 55, 473–481 (2019). https://doi.org/10.1007/s11626-019-00367-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-019-00367-y

Keywords

Search

Navigation