Microfabricated multilayer parylene-C stencils for the generation of patterned dynamic co-cultures.
Co-culturing different cell types can be useful to engineer a more in vivo-like microenvironment for cells in culture. Recent approaches to generating cellular co-cultures have used microfabrication technologies to regulate the degree of cell-cell contact between different cell types. However, these approaches are often limited to the co-culture of only two cell types in static cultures. The dynamic aspect of cell-cell interaction, however, is a key regulator of many biological processes such as early development, stem cell differentiation, and tissue regeneration. In this study, we describe a micropatterning technique based on microfabricated multilayer parylene-C stencils and demonstrate the potential of parylene-C technology for co-patterning of proteins and cells with the ability to generate a series of at least five temporally controlled patterned co-cultures. We generated dynamic co-cultures of murine embryonic stem cells in culture with various secondary cell types that could be sequentially introduced and removed from the co-cultures. Our studies suggested that dynamic co-cultures generated by using parylene-C stencils may be applicable in studies investigating cellular interactions in controlled microenvironments such as studies of ES cell differentiation, wound healing and development.
Jinno
Satoshi
S
Department of Medicine, Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Moeller
Hannes-Christian
HC
Chen
Chia-Ling
CL
Rajalingam
Bimal
B
Chung
Bong Geun
BG
Dokmeci
Mehmet R
MR
Khademhosseini
Ali
A
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000295:instrumentation* / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology; D051379:Mice; D008855:Microscopy, Electron, Scanning; D041681:NIH 3T3 Cells; D011108:Polymers / Q000737:chemistry*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D014945:Wound Healing; D014992:Xylenes / Q000737:chemistry*
D001672:Biocompatible Materials; D011108:Polymers; D014992:Xylenes; C011055:parylene