Gelatin-based laser direct-write technique for the precise spatial patterning of cells.
Laser direct-writing provides a method to pattern living cells in vitro, to study various cell-cell interactions, and to build cellular constructs. However, the materials typically used may limit its long-term application. By utilizing gelatin coatings on the print ribbon and growth surface, we developed a new approach for laser cell printing that overcomes the limitations of Matrigel™. Gelatin is free of growth factors and extraneous matrix components that may interfere with cellular processes under investigation. Gelatin-based laser direct-write was able to successfully pattern human dermal fibroblasts with high post-transfer viability (91% ± 3%) and no observed double-strand DNA damage. As seen with atomic force microscopy, gelatin offers a unique benefit in that it is present temporarily to allow cell transfer, but melts and is removed with incubation to reveal the desired application-specific growth surface. This provides unobstructed cellular growth after printing. Monitoring cell location after transfer, we show that melting and removal of gelatin does not affect cellular placement; cells maintained registry within 5.6 ± 2.5 μm to the initial pattern. This study demonstrates the effectiveness of gelatin in laser direct-writing to create spatially precise cell patterns with the potential for applications in tissue engineering, stem cell, and cancer research.
Schiele
Nathan R
NR
Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, New York, USA.
Chrisey
Douglas B
DB
Corr
David T
DT
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D048430:Cell Shape; D002470:Cell Survival; D004249:DNA Damage; D020405:Dermis / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis; D005455:Fluorescent Antibody Technique; D005780:Gelatin / Q000737:chemistry*; D006657:Histones / Q000378:metabolism; D006801:Humans; D007834:Lasers; D018625:Microscopy, Atomic Force; D010766:Phosphorylation; D034421:Sus scrofa; D013997:Time Factors
D005353:Fibronectins; C468783:H2AX protein, human; D006657:Histones; D005780:Gelatin