Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing.
BACKGROUND
Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale.


RESULTS
Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms.


CONCLUSIONS
Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology.
Liebmann
Thomas
T
Cell Physics, Department of Applied Physics, Royal Institute of Technology, S-106 91 Stockholm, Sweden. liebmann@kth.se
Rydholm
Susanna
S
Akpe
Victor
V
Brismar
Hjalmar
H
D000596:Amino Acids; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D019556:COS Cells; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002470:Cell Survival; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004151:Dipeptides; D004285:Dogs; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005449:Fluorenes; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D008422:Materials Testing; D010649:Phenylalanine / Q000031:analogs & derivatives; D051381:Rats; D046509:Tissue Culture Techniques / Q000295:instrumentation / Q000379:methods; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D054457:Tissue Scaffolds
D000596:Amino Acids; D004151:Dipeptides; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; C026650:phenylalanylphenylalanine; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D010649:Phenylalanine