Using growth factor arrays and micropatterned co-cultures to induce hepatic differentiation of embryonic stem cells

Nazgul Tuleuova, Ji Youn Lee, Jennifer Lee, Erlan Ramanculov, Mark A. Zern, Alexander Revzin

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

The success in driving embryonic stem cells towards hepatic lineage has been confounded by the complexity and cost of differentiation protocols that employ large quantities of expensive growth factors (GFs). Instead of supplementing culture media with soluble GFs, we investigated cultivation and differentiation of mouse embryonic stem cells (mESCs) on printed arrays of GFs. Hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF) and bone morphogenetic protein (BMP4) were mixed in solution with fibronectin and collagen (I) and then printed onto silane-modified glass slides to form 500 μm diameter protein spots. mESCs were cultured on top of GF spots for up to 12 days and analyzed by RT-PCR and immunostaining at different time points. The stem cells residing on HGF-containing combinations of GFs exhibited requisite features of hepatic differentiation including pronounced loss in pluripotency (Oct4), transient (up and down) expression of endoderm (Sox17) and upregulation of early hepatic markers - albumin and alpha-fetoprotein. The hepatic differentiation was enhanced further by adding hepatic stellate cells to surfaces that already contained mESCs on GF spots. A combination of co-culture with non-parenchymal liver cells and the optimal GF stimulation was found to induce endoderm and hepatic phenotype earlier and to a much greater extent than the GF arrays or micropatterned co-cultures used individually. While this paper investigated hepatic differentiation of mouse ESCs, our findings and stem cell culture approaches are likely to be relevant for human ESC cultivation. Overall, the platform combining printed GF arrays and heterotypic co-cultures will be broadly applicable for identifying the composition of the microenvironment niche for ESC differentiation into various tissue types.

Original languageEnglish
Pages (from-to)9221-9231
Number of pages11
JournalBiomaterials
Volume31
Issue number35
DOIs
Publication statusPublished - Dec 2010
Externally publishedYes

Fingerprint

Embryonic Stem Cells
Coculture Techniques
Stem cells
Cell culture
Intercellular Signaling Peptides and Proteins
Liver
Hepatocyte Growth Factor
Endoderm
Stem Cells
Silanes
Bone Morphogenetic Proteins
Hepatic Stellate Cells
alpha-Fetoproteins
Fibroblast Growth Factor 2
Fibronectins
Proteins
Culture Media
Glass
Collagen
Albumins

Keywords

  • Embryonic stem cells
  • Growth factor arrays
  • Hepatic differentiation
  • Micropatterned co-cultures

ASJC Scopus subject areas

  • Biomaterials
  • Bioengineering
  • Ceramics and Composites
  • Mechanics of Materials
  • Biophysics

Cite this

Using growth factor arrays and micropatterned co-cultures to induce hepatic differentiation of embryonic stem cells. / Tuleuova, Nazgul; Lee, Ji Youn; Lee, Jennifer; Ramanculov, Erlan; Zern, Mark A.; Revzin, Alexander.

In: Biomaterials, Vol. 31, No. 35, 12.2010, p. 9221-9231.

Research output: Contribution to journalArticle

Tuleuova, Nazgul ; Lee, Ji Youn ; Lee, Jennifer ; Ramanculov, Erlan ; Zern, Mark A. ; Revzin, Alexander. / Using growth factor arrays and micropatterned co-cultures to induce hepatic differentiation of embryonic stem cells. In: Biomaterials. 2010 ; Vol. 31, No. 35. pp. 9221-9231.
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