The effects of soluble growth factors on embryonic stem cell differentiation inside of fibrin scaffolds.
The goal of this research was to determine the effects of different growth factors on the survival and differentiation of murine embryonic stem cell-derived neural progenitor cells (ESNPCs) seeded inside of fibrin scaffolds. Embryoid bodies were cultured for 8 days in suspension, retinoic acid was applied for the final 4 days to induce ESNPC formation, and then the EBs were seeded inside of three-dimensional fibrin scaffolds. Scaffolds were cultured in the presence of media containing different doses of the following growth factors: neurotrophin-3 (NT-3), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF)-AA, ciliary neurotrophic factor, and sonic hedgehog (Shh). The cell phenotypes were characterized using fluorescence-activated cell sorting and immunohistochemistry after 14 days of culture. Cell viability was also assessed at this time point. Shh (10 ng/ml) and NT-3 (25 ng/ml) produced the largest fractions of neurons and oligodendrocytes, whereas PDGF (2 and 10 ng/ml) and bFGF (10 ng/ml) produced an increase in cell viability after 14 days of culture. Combinations of growth factors were tested based on the results of the individual growth factor studies to determine their effect on cell differentiation. The incorporation of ESNPCs and growth factors into fibrin scaffolds may serve as potential treatment for spinal cord injury.
Willerth
Stephanie M
SM
Department of Biomedical Engineering, Washington University, Campus Box 1097, One Brookings Drive, St. Louis, Missouri 63130, USA.
Faxel
Tracy E
TE
Gottlieb
David I
DI
Sakiyama-Elbert
Shelly E
SE
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D020934:Ciliary Neurotrophic Factor / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism*; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D053823:Hedgehog Proteins / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D051379:Mice; D008954:Models, Biological; D009414:Nerve Growth Factors / Q000494:pharmacology; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D017978:Receptors, Growth Factor / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D020934:Ciliary Neurotrophic Factor; D004338:Drug Combinations; D053823:Hedgehog Proteins; D036341:Intercellular Signaling Peptides and Proteins; D009414:Nerve Growth Factors; D010982:Platelet-Derived Growth Factor; D017978:Receptors, Growth Factor; C465570:Shh protein, mouse; C518911:neurotropin 3, mouse; D005346:Fibroblast Growth Factors; D005337:Fibrin