Laser Direct-Write of Embryonic Stem Cells and Cells Encapsulated in Alginate Beads for Engineered Biological Constructs
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Laser Direct-Write of Embryonic Stem Cells and Cells Encapsulated in Alginate Beads for Engineered Biological Constructs T.B. Phamduy1, A.D. Dias1, N. Abdul Raof2, N.R. Schiele1, D.T. Corr1, Y. Xie2, and D.B. Chrisey1,3 1
Biomedical Engineering Department, Rensselaer Polytechnic Institute, 110 Eighth St., Troy, NY 12180 College of Nanoscale Science and Engineering, University at Albany, State University of New York, 257 Fuller Rd., Albany, NY 12203 Material Science and Engineering, Rensselaer Polytechnic Institute, 110 Eighth St., Troy, NY 12180
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Abstract The ability to control the deposition of mouse embryonic stem cells (mESCs), and mESCs encapsulated in 200-μm diameter alginate microbeads, into customized patterns has recently been achieved using laser direct-write (LDW). Gelatin-based LDW was utilized to target and reproducibly deposit groups of cells directly onto receiving substrate surfaces. Live/dead staining for cell viability and immunocytochemistry for the pluripotency marker, Oct-4, indicated that transferred mESCs were viable following transfer, and maintained an important embryonic stem cell marker, respectively. LDW was further used to print mESCs encapsulated in hydrogel microbeads into customized patterns on a single-bead basis. Recent efforts have also achieved patterns of discrete co-cultures of mESCs and breast cancer cells in separate hydrogel microbeads. Altogether, we demonstrated the feasibility of LDW to print patterns of mESCs and mESC-microbeads for the biomimetic assembly of engineered cellular constructs and tissue models.
Introduction Stem cells play a pivotal role in the future of regenerative medicine and are characterized by their dual ability to self-renew or differentiate into specialized cell types that give rise to diverse tissues. 1–3 Recapitulating the in vivo microenvironment through precise in vitro biofabrication may control stem cell fate and facilitate engineered tissue development. 2,4 The inspiration for cellular programming comes from the niche, an anatomical microenvironment that subjects stem cells to a multitude of disparate biological stimuli (e.g., cell-cell interactions, soluble factors, biomechanical cues) 1. In vivo cell growth, differentiation, and tissue development involve a complex combination and dynamic interplay of many cues that are difficult to reproduce with current tissue engineering technologies. Thus, there exists a demand for novel approaches to biofabrication 4. To fully realize the potential clinical applications of stem cells and drive their differentiation towards a desired fate, novel techniques must be able to isolate and re-integrate the environmental cues to produce artificial tissue. In particular, the ability to spatially control cellular assembly, and concomitantly the interactions between neighboring homogeneous and heterogeneous cell types, influences the mode of cell-cell communication (i.e., direct cell contact vs. paracrine signaling) and stem cell differentiation 5. Idealized tissue from spatially-resolved cell cultures must
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