Skip to main content
Log in

Initial cell adhesion of three cell types in the presence and absence of serum proteins

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

With the development of a wide range of new biomaterials for the sensing of different cell behaviour, it is important to consider whether the cells tested in vitro are in direct contact with the material or whether cell–biomaterial contact is mediated by an interfacial layer of proteins originating from the culture medium or from the cells themselves. Thus, this study describes the differences between the cell adhesion mediated by proteins originating from foetal bovine serum and without the presence of such proteins 2 h following cell seeding exemplarily with different cell types (an osteoblastic cell line, primary fibroblasts, and mesenchymal stem cells). Three of the examined cell types were found to react differently to differing conditions in terms of cell shape, area, and number. Nevertheless, the expression and localization of the various proteins involved in cell adhesion and signalling (CD44, vinculin, talin, actin, focal adhesion kinase, Rho-GTPases and extracellular signal-regulated kinases 1 and 2) were, in general, similar with respect to all the cell types tested, albeit varying according to the presence or absence of serum. Moreover, no classical focal adhesions were formed during cell adhesion without serum proteins, while different signalling pathways were involved in this process. The study systematically describes and discusses the cell adhesion of three different human cell types to a well-known substrate without the presence of external proteins and it is hoped that this knowledge will be subsequently applied in biomaterial applications in which the presence of external proteins is undesirable (e.g. for biosensing purposes).

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Abumaree MH, Al Jumah MA, Kalionis B, Jawdat D, Al Khaldi A, AlTalabani AA, Knawy BA (2013) Phenotypic and functional characterization of mesenchymal stem cells from chorionic villi of human term placenta. Stem Cell Rev Rep 9:16–31

    Article  CAS  Google Scholar 

  • Anselme K, Ploux L, Ponche A (2010) Cell/material interfaces: influence of surface chemistry and surface topography on cell adhesion. J Adhes Sci Technol 24:831–852

    Article  CAS  Google Scholar 

  • Ashida N, Takechi H, Kita T, Arai H (2003) Vortex-mediated mechanical stress induces integrin-dependent cell adhesion mediated by inositol 1,4,5-trisphosphate-sensitive Ca2 + release in THP-1 cells. J Biol Chem 278:9327–9331

    Article  CAS  PubMed  Google Scholar 

  • Audiffred JF, De Leo SE, Brown PK, Hale-Donze H, Monroe WT (2010) Characterization and applications of serum-free induced adhesion in jurkat suspension cells. Biotechnol Bioeng 106:784–793

    Article  CAS  PubMed  Google Scholar 

  • Calderwood DA, Campbell ID, Critchley DR (2013) Talins and kindlins: partners in integrin-mediated adhesion. Nat Rev Mol Cell Biol 14:503–517

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Carre A, Lacarriere V (2010) How substrate properties control cell adhesion. a physical–chemical approach. J Adhes Sci Technol 24:815–830

    Article  CAS  Google Scholar 

  • Chen RH, Sarnecki C, Blenis J (1992) Nuclear localization and regulation of erk- and rsk-encoded protein kinases. Mol Cell Biol 12:915–927

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chiarugi P, Giannoni E (2008) Anoikis: a necessary death program for anchorage-dependent cells. Biochem Pharmacol 76:1352–1364

    Article  CAS  PubMed  Google Scholar 

  • Crisp M, Liu Q, Roux K, Rattner JB, Shanahan C, Burke B, Stahl PD, Hodzic D (2006) Coupling of the nucleus and cytoplasm: role of the LINC complex. J Cell Biol 172:41–53

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Critchley DR (2009) Biochemical and structural properties of the integrin-associated cytoskeletal protein talin. Annu Rev Biophys 38:235–254

    Article  CAS  PubMed  Google Scholar 

  • Frisch SM, Francis H (1994) Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol 124:619–626

    Article  CAS  PubMed  Google Scholar 

  • Geiger B, Yamada KM (2011) Molecular architecture and function of matrix adhesions. Csh Perspect Biol 3

  • Humphries MJ, Akiyama SK, Komoriya A, Olden K, Yamada KM (1986) Identification of an alternatively spliced site in human plasma fibronectin that mediates cell type-specific adhesion. J Cell Biol 103:2637–2647

    Article  CAS  PubMed  Google Scholar 

  • Humphries JD, Wang P, Streuli C, Geiger B, Humphries MJ, Ballestrem C (2007) Vinculin controls focal adhesion formation by direct interactions with talin and actin. J Cell Biol 179:1043–1057

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jalkanen S, Jalkanen M (1992) Lymphocyte CD44 binds the COOH-terminal heparin-binding domain of fibronectin. J Cell Biol 116:817–825

    Article  CAS  PubMed  Google Scholar 

  • Kalbacova M, Broz A, Kromka A, Babchenko O, Kalbac M (2011) Controlled oxygen plasma treatment of single-walled carbon nanotube films improves osteoblastic cells attachment and enhances their proliferation. Carbon 49:2926–2934

    Article  CAS  Google Scholar 

  • Kanchanawong P, Shtengel G, Pasapera AM, Ramko EB, Davidson MW, Hess HF, Waterman CM (2010) Nanoscale architecture of integrin-based cell adhesions. Nature 468:580–584

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Khalili AA, Ahmad MR (2015) A review of cell adhesion studies for biomedical and biological applications. Int J Mol Sci 16:18149–18184

    Article  CAS  PubMed  Google Scholar 

  • Koblinski JE, Wu M, Demeler B, Jacob K, Kleinman HK (2005) Matrix cell adhesion activation by non-adhesion proteins. J Cell Sci 118:2965–2974

    Article  CAS  PubMed  Google Scholar 

  • Kolyada AY, Riley KN, Herman IM (2003) Rho GTPase signaling modulates cell shape and contractile phenotype in an isoactin-specific manner. Am J Physiol-Cell Ph 285:C1116–C1121

    Article  CAS  Google Scholar 

  • Krebs HA (1950) Chemical composition of blood plasma and serum. Annu Rev Biochem 19:409–430

    Article  CAS  PubMed  Google Scholar 

  • Mager MD, LaPointe V, Stevens MM (2011) Exploring and exploiting chemistry at the cell surface. Nat Chem 3:582–589

    Article  CAS  PubMed  Google Scholar 

  • Ostrovska L, Broz A, Fucikova A, Belinova T, Sugimoto H, Kanno T, Fujii M, Valenta J, Kalbacova MH (2016) The impact of doped silicon quantum dots on human osteoblasts. Rsc Adv 6:63403–63413

    Article  CAS  Google Scholar 

  • Pautke C, Schieker M, Tischer T, Kolk A, Neth P, Mutschler W, Milz S (2004) Characterization of osteosarcoma cell lines MG-63, Saos-2 and U-2 OS in comparison to human osteoblasts. Anticancer Res 24:3743–3748

    CAS  PubMed  Google Scholar 

  • Ponta H, Sherman L, Herrlich PA (2003) CD44: from adhesion molecules to signalling regulators. Nat Rev Mol Cell Biol 4:33–45

    Article  CAS  PubMed  Google Scholar 

  • Rodan SB, Imai Y, Thiede MA, Wesolowski G, Thompson D, Barshavit Z, Shull S, Mann K, Rodan GA (1987) Characterization of a human osteosarcoma cell-line (Saos-2) with osteoblastic properties. Cancer Res 47:4961–4966

    CAS  PubMed  Google Scholar 

  • Roskoski R Jr (2012) ERK1/2 MAP kinases: structure, function, and regulation. Pharmacol Res Off J Ital Pharmacol Soc 66:105–143

    CAS  Google Scholar 

  • Ruoslahti E (1996) RGD and other recognition sequences for integrins. Annu Rev Cell Dev Biol 12:697–715

    Article  CAS  PubMed  Google Scholar 

  • Ruszova E, Cheel J, Pavek S, Moravcova M, Hermannova M, Matejkova I, Spilkova J, Velebny V, Kubala L (2013) Epilobium angustifolium extract demonstrates multiple effects on dermal fibroblasts in vitro and skin photo-protection in vivo. Gen Physiol Biophys 32:347–359

    Article  CAS  PubMed  Google Scholar 

  • Ryzhkov P, Prass M, Gummich M, Kuhn JS, Oettmeier C, Dobereiner HG (2010) Adhesion patterns in early cell spreading. J Phys Condens Matter Inst Phys J 22:194106

    Article  Google Scholar 

  • Sailem H, Bousgouni V, Cooper S, Bakal C (2014) Cross-talk between Rho and Rac GTPases drives deterministic exploration of cellular shape space and morphological heterogeneity. Open Biol 4

  • Saoncella S, Echtermeyer F, Denhez F, Nowlen JK, Mosher DF, Robinson SD, Hynes RO, Goetinck PF (1999) Syndecan-4 signals cooperatively with integrins in a Rho-dependent manner in the assembly of focal adhesions and actin stress fibers. Proc Natl Acad Sci USA 96:2805–2810

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schlaepfer DD, Hauck CR, Sieg DJ (1999) Signaling through focal adhesion kinase. Prog Biophys Mol Biol 71:435–478

    Article  CAS  PubMed  Google Scholar 

  • Sundin M, Ringden O, Sundberg B, Nava S, Gotherstrom C, Le Blanc K (2007) No alloantibodies against mesenchymal stromal cells, but presence of anti-fetal calf serum antibodies, after transplantation in allogeneic hematopoietic stem cell recipients. Haematologica 92:1208–1215

    Article  CAS  PubMed  Google Scholar 

  • Verdanova M, Rezek B, Broz A, Ukraintsev E, Babchenko O, Artemenko A, Izak T, Kromka A, Kalbac M, Kalbacova MH (2016) Nanocarbon allotropes—graphene and nanocrystalline diamond—promote cell proliferation. Small 12:2499–2509

    Article  CAS  PubMed  Google Scholar 

  • Wei JH, Igarashi T, Okumori N, Igarashi T, Maetani T, Liu BL, Yoshinari M (2009) Influence of surface wettability on competitive protein adsorption and initial attachment of osteoblasts. Biomed Mater 4

  • Wilson CJ, Clegg RE, Leavesley DI, Pearcy MJ (2005) Mediation of biomaterial-cell interactions by adsorbed proteins: a review. Tissue Eng 11:1–18

    Article  CAS  PubMed  Google Scholar 

  • Yoon S, Seger R (2006) The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions. Growth Factors 24:21–44

    Article  CAS  PubMed  Google Scholar 

  • Zaidel-Bar R, Cohen M, Addadi L, Geiger B (2004) Hierarchical assembly of cell-matrix adhesion complexes. Biochem Soc Trans 32:416–420

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by: Charles University in Prague, First Faculty of Medicine: project PROGRES Q26, Faculty of Medicine in Pilsen: project SVV 260 390 and GAUK 400215; further by the National Sustainability Program I (NPU I) Nr. LO1503 provided by the Ministry of Education, Youth and Sports of the Czech Republic and project 15-25813A-AZV CR. Special thanks go to Blanka Bilkova for her technical assistance.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Martina Verdanova.

Ethics declarations

Conflicts of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Verdanova, M., Sauerova, P., Hempel, U. et al. Initial cell adhesion of three cell types in the presence and absence of serum proteins. Histochem Cell Biol 148, 273–288 (2017). https://doi.org/10.1007/s00418-017-1571-7

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-017-1571-7

Keywords

Navigation