Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Oncology Letters
      • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Biomedical Reports
      • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • Information for Authors
    • Information for Reviewers
    • Information for Librarians
    • Information for Advertisers
    • Conferences
  • Language Editing
Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • For Authors
    • For Reviewers
    • For Librarians
    • For Advertisers
    • Conferences
  • Language Editing
Login Register Submit
  • This site uses cookies
  • You can change your cookie settings at any time by following the instructions in our Cookie Policy. To find out more, you may read our Privacy Policy.

    I agree
Search articles by DOI, keyword, author or affiliation
Search
Advanced Search
presentation
Experimental and Therapeutic Medicine
Join Editorial Board Propose a Special Issue
Print ISSN: 1792-0981 Online ISSN: 1792-1015
Journal Cover
June-2016 Volume 11 Issue 6

Full Size Image

Sign up for eToc alerts
Recommend to Library

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

Medicine International

An International Open Access Journal Devoted to General Medicine.

Journal Cover
June-2016 Volume 11 Issue 6

Full Size Image

Sign up for eToc alerts
Recommend to Library

  • Article
  • Citations
    • Cite This Article
    • Download Citation
    • Create Citation Alert
    • Remove Citation Alert
    • Cited By
  • Similar Articles
    • Related Articles (in Spandidos Publications)
    • Similar Articles (Google Scholar)
    • Similar Articles (PubMed)
  • Download PDF
  • Download XML
  • View XML
Article

Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells

  • Authors:
    • Hua Yang
    • Ying Qiu
    • Xianghui Zeng
    • Yan Ding
    • Jianye Zeng
    • Kehuan Lu
    • Dongsheng Li
  • View Affiliations / Copyright

    Affiliations: Reproductive Medical Center of Nanning Second People's Hospital, Third Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530031, P.R. China, Life Science Research Institute, Taihe Hospital, Shiyan, Hubei 442000, P.R. China, State Key Laboratory for Conservation and Utilization of Subtropical Agro‑Bioresources, Animal Reproduction Institute, Guangxi University, Nanning, Guangxi 530004, P.R. China
  • Pages: 2321-2328
    |
    Published online on: March 30, 2016
       https://doi.org/10.3892/etm.2016.3204
  • Expand metrics +
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
Cited By (CrossRef): 0 citations Loading Articles...

This article is mentioned in:



Abstract

The aim of the present study was to investigate the effects of feeder layers composed of various ratios of mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (hFFs) on the growth of human embryonic stem cells (hESCs). In addition, the secretion levels of basic fibroblast growth factor (bFGF) by the feeder layers was detected. MEFs and hFFs were treated with mitomycin C and seeded onto gelatin‑coated plates at a density of 1x108 cells/l. The hFFs and MEFs were combined and plated at the following ratios: 0:1, 1:2, 1:1, 2:1 and 1:0. The secretion of bFGF by the various hFF/MEF ratio feeder layers was detected using an enzyme‑linked immunosorbent assay. Subsequently, hESCs were cultured on top of the various feeder layers. The differences in the cellular morphology of the hESCs were observed using microscopy, and the expression levels alkaline phosphatase (AKP) and octamer‑binding transcription factor 4 (OCT‑4) were detected using immunohistochemical analysis as indicators of differentiation status. The results showed that the hFFs secreted substantial quantities of bFGF, while no bFGF was secreted by the MEFs. The clones of hESC growing on the feeder layer containing MEF or hFF alone were flat. By contrast, hESC clones grown on a mixed feeder layer containing hFFs + MEFs at a ratio of 1:1 exhibited an accumulated growth with a clear edge, as compared with the other ratios. In addition, hESCs growing on the feeder layer were positive for the expression of AKP and OCT‑4. In summary, feeder layer hFFs secreted bFGF, while MEFs did not, indicating that bFGF is not the only factor that supports the growth and differentiation of hESCs. The optimal growth of hESCs was achieved using a mixed feeder layer composed of hFFs + MEFs at a ratio of 1:1.
View Figures

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

View References

1 

Whyte M, Hubbard R, Meliconi R, Whidborne M, Eaton V, Bingle C, Timms J, Duff G, Facchini A, Pacilli A, et al: Increased risk of fibrosing alveolitis associated with interleukin-1 receptor antagonist and tumor necrosis factor-alpha gene polymorphisms. Am J Respir Crit Care Med. 162:755–758. 2000. View Article : Google Scholar : PubMed/NCBI

2 

Bjorklund LM, Sánchez-Pernaute S, Chung S, Andersson T, Chen IY, McNaught KS, Brownell AL, Jenkins BG, Wahlestedt C, Kim KS and Isacson O: Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model. Proc Natl Acad Sci USA. 99:2244–2349. 2002. View Article : Google Scholar

3 

Brüstle O, Jones KN, Learish RD, Karram K, Choudhary K, Wiestler OD, Duncan ID and McKay RD: Embryonic stem cell-derived glial precursors: A source of myelinating transplants. Science. 285:754–756. 1999. View Article : Google Scholar : PubMed/NCBI

4 

Liu S, Qu Y, Stewart TJ, Howard MJ, Chakrabortty S, Holekamp TF and McDonald JW: Embryonic stem cells differentiate into oligodendrocytes and myelinate in culture and after spinal cord transplantation. Proc Natl Acad Sci USA. 97:6126–6131. 2000. View Article : Google Scholar : PubMed/NCBI

5 

Amit M, Margulets V, Segev H, Shariki K, Laevsky I, Coleman R and Itskovitz-Eldor J: Human feeder layers for human embryonic stem cells. Biol Reprod. 68:2150–2156. 2003. View Article : Google Scholar : PubMed/NCBI

6 

Hovatta O, Mikkola M, Gertow K, Strömberg AM, Inzunza J, Hreinsson J, Rozell B, Blennow E, Andäng M and Ahrlund-Richter L: A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells. Hum Reprod. 18:1404–1409. 2003. View Article : Google Scholar : PubMed/NCBI

7 

Inzunza J, Gertow K, Strömberg MA, Matilainen E, Blennow E, Skottman H, Wolbank S, Ahrlund-Richter L and Hovatta O: Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells. Stem Cells. 23:544–549. 2005. View Article : Google Scholar : PubMed/NCBI

8 

Nieto A, Cabrera CM, Catalina P, Cobo F, Barnie A, Cortés JL, del Barroso Jesus A, Montes R and Concha A: Effect of mitomycin-C on human foreskin fibroblasts used as feeders in human embryonic stem cells: Immunocytochemistry MIBI score and DNA ploidy and apoptosis evaluated by flow cytometry. Cell Biol Int. 31:269–278. 2007. View Article : Google Scholar : PubMed/NCBI

9 

Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS and Jones JM: Embryonic stem cell lines derived from human blastocytes. Science. 282:1145–1147. 1998. View Article : Google Scholar : PubMed/NCBI

10 

Troy TC, Rahbar R, Diker B and Turksen K: Immunolocalization in the epidermis. Methods Mol Biol. 289:113–120. 2005.PubMed/NCBI

11 

Saxena S, Hanwate M, Deb K, Sharma V and Totey S: FGF2 secreting human fibroblast feeder cells: A novel culture system for human embryonic stem cells. Mol Reprod Dev. 75:1523–1532. 2008. View Article : Google Scholar : PubMed/NCBI

12 

Gavrilov S, Marolt D, Douglas NC, Prosser RW, Khalid I, Sauer MV, Landry DW, Vunjak-Novakovic G and Papaioannou VE: Derivation of two new human embryonic stem cell lines from nonviable human embryos. Stem Cells Int. 2011:7653782011. View Article : Google Scholar : PubMed/NCBI

13 

Vaajasaari H, Ilmarinen T, Juuti-Uusitalo K, Rajala K, Onnela N, Narkilahti S, Suuronen R, Hyttinen J, Uusitalo H and Skottman H: Toward the defined and xeno-free differentiation of functional human pluripotent stem cell-derived retinal pigment epithelial cells. Mol Vis. 17:558–575. 2011.PubMed/NCBI

14 

Ding Y, Yang H, Yu L, Xu CL, Zeng Y, Qiu Y and Li DS: Feeder-free and xeno-free culture of human pluripotent stem cells using UCBS matrix. Cell Biol Int. 39:1111–1119. 2015. View Article : Google Scholar : PubMed/NCBI

15 

Coulombe PA and Wong I: Cytoplasmic intermediate filaments revealed as dynamic and multipurpose scaffolds. Nat Cell Biol. 6:699–706. 2004. View Article : Google Scholar : PubMed/NCBI

16 

Xu C, Inokuma MS, Denham J, Golds K, Kundu P, Gold JD and Carpenter MK: Feeder-free growth of undifferentiated human embryonic stem cells. Nat Biotechnol. 19:971–974. 2001. View Article : Google Scholar : PubMed/NCBI

17 

Hirai H, Karian P and Kikyo N: Regulation of embryonic stem cell self-renewal and pluripotency by leukaemia inhibitory factor. Biochem J. 438:11–23. 2011. View Article : Google Scholar : PubMed/NCBI

18 

Salli U, Fox TE, Carkaci-Salli N, Sharma A, Robertson GP, Kester M and Vrana KE: Propagation of undifferentiated human embryonic stem cells with nano-liposomal ceramide. Stem Cells Dev. 18:55–65. 2009. View Article : Google Scholar : PubMed/NCBI

19 

Yabut O and Bernstein HS: The promise of human embryonic stem cells in aging-associated diseases. Aging (Albany NY). 3:494–508. 2011. View Article : Google Scholar : PubMed/NCBI

20 

Huang YC, Wang TW, Sun JS and Lin FH: Investigation of mitomycin-C-treated fibroblasts in 3-D collagen gel and conditioned medium for keratinocyte proliferation. Artif Organs. 30:150–159. 2006. View Article : Google Scholar : PubMed/NCBI

21 

Zhou YP, Rochat A, Hatzfeld A, Peiffer I, Barbet R, Hatzfeld J and Li ML: bFGF-stimulated MEF-conditioned medium is capable of maintaining human embryonic stem cells. Fen Zi Xi Bao Sheng Wu Xue Bao. 42:193–199. 2009.(In Chinese). PubMed/NCBI

22 

Xu HF and Suming Z: Feeder-free growth of human embryonic stem cells supported by basic fibroblast growth factor. Zhong Guo Zu Zhi Gong Cheng Yan Jiu Yu Lin Chuang Kang Fu. 14:1111–1114. 2010.(In Chinese).

23 

Chiba S, Lee YM, Zhou W and Freed CR: Noggin enhances dopamine neuron production from human embryonic stem cells and improves behavioral outcome after transplantation into Parkinsonian rats. Stem Cells. 26:2810–2120. 2008. View Article : Google Scholar : PubMed/NCBI

24 

Eiselleova L, Peterkova I, Neradil J, Slaninova I, Hampl A and Dvorak P: Comparative study of mouse and human feeder cells for human embryonic stem cells. Int J Dev Biol. 52:353–363. 2008. View Article : Google Scholar : PubMed/NCBI

25 

Li SS, Liu YH, Tseng CN, Chung TL, Lee TY and Singh S: Characterization and gene expression profiling of five new human embryonic stem cell lines derived in Taiwan. Stem Cells Dev. 15:532–555. 2006. View Article : Google Scholar : PubMed/NCBI

26 

Mandal A, Tipnis S, Pal R, Ravindran G, Bose B, Patki A, Rao MS and Khanna A: Characterization and in vitro differentiation potential of a new human embryonic stem cell line, ReliCellhES1. Differentiation. 74:81–90. 2006. View Article : Google Scholar : PubMed/NCBI

27 

Lee JB, Lee JE, Park JH, Kim SJ, Kim MK, Roh SI and Yoon HS: Establishment and maintenance of human embryonic stem cell lines on human feeder cells derived from uterine endometrium under serum-free condition. Biol Reprod. 72:42–49. 2005. View Article : Google Scholar : PubMed/NCBI

28 

Genbacev O, Krtolica A, Zdravkovic T, Brunette E, Powell S, Nath A, Caceres E, McMaster M, McDonagh S, Li Y, et al: Serum-free derivation of human embryonic stem cell lines on human placental fibroblast feeders. Fertil Steril. 83:1517–1529. 2005. View Article : Google Scholar : PubMed/NCBI

29 

Richards M, Tan S, Fong CY, Biswas A, Chan WK and Bongso A: Comparative evaluation of various human feeders for prolonged undifferentiated growth of human embryonic stem cells. Stem Cells. 21:546–556. 2003. View Article : Google Scholar : PubMed/NCBI

30 

Xu C, Jiang J, Sottile V, McWhir J, Lebkowski J and Carpenter MK: Immortalized fibroblast-like cells derived from human embryonic stem cells support undifferentiated cell growth. Stem Cells. 22:972–980. 2004. View Article : Google Scholar : PubMed/NCBI

31 

Wang Q, Fang ZF, Jin F, Lu Y, Gai H and Sheng HZ: Derivation and growing human embryonic stem cells on feeders derived from themselves. Stem Cells. 23:1221–1227. 2005. View Article : Google Scholar : PubMed/NCBI

32 

Stojkovic P, Lako M, Stewart R, Przyborski S, Armstrong L, Evans J, Murdoch A, Strachan T and Stojkovic M: An autogeneic feeder cell system that efficiently supports growth of undifferentiated human embryonic stem cells. Stem Cells. 23:306–314. 2005. View Article : Google Scholar : PubMed/NCBI

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Yang H, Qiu Y, Zeng X, Ding Y, Zeng J, Lu K and Li D: Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells. Exp Ther Med 11: 2321-2328, 2016.
APA
Yang, H., Qiu, Y., Zeng, X., Ding, Y., Zeng, J., Lu, K., & Li, D. (2016). Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells. Experimental and Therapeutic Medicine, 11, 2321-2328. https://doi.org/10.3892/etm.2016.3204
MLA
Yang, H., Qiu, Y., Zeng, X., Ding, Y., Zeng, J., Lu, K., Li, D."Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells". Experimental and Therapeutic Medicine 11.6 (2016): 2321-2328.
Chicago
Yang, H., Qiu, Y., Zeng, X., Ding, Y., Zeng, J., Lu, K., Li, D."Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells". Experimental and Therapeutic Medicine 11, no. 6 (2016): 2321-2328. https://doi.org/10.3892/etm.2016.3204
Copy and paste a formatted citation
x
Spandidos Publications style
Yang H, Qiu Y, Zeng X, Ding Y, Zeng J, Lu K and Li D: Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells. Exp Ther Med 11: 2321-2328, 2016.
APA
Yang, H., Qiu, Y., Zeng, X., Ding, Y., Zeng, J., Lu, K., & Li, D. (2016). Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells. Experimental and Therapeutic Medicine, 11, 2321-2328. https://doi.org/10.3892/etm.2016.3204
MLA
Yang, H., Qiu, Y., Zeng, X., Ding, Y., Zeng, J., Lu, K., Li, D."Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells". Experimental and Therapeutic Medicine 11.6 (2016): 2321-2328.
Chicago
Yang, H., Qiu, Y., Zeng, X., Ding, Y., Zeng, J., Lu, K., Li, D."Effect of a feeder layer composed of mouse embryonic and human foreskin fibroblasts on the proliferation of human embryonic stem cells". Experimental and Therapeutic Medicine 11, no. 6 (2016): 2321-2328. https://doi.org/10.3892/etm.2016.3204
Follow us
  • Twitter
  • LinkedIn
  • Facebook
About
  • Spandidos Publications
  • Careers
  • Cookie Policy
  • Privacy Policy
How can we help?
  • Help
  • Live Chat
  • Contact
  • Email to our Support Team