A combinatorial cell-laden gel microarray for inducing osteogenic differentiation of human mesenchymal stem cells.
Development of three dimensional (3D) microenvironments that direct stem cell differentiation into functional cell types remains a major challenge in the field of regenerative medicine. Here, we describe a new platform to address this challenge by utilizing a robotic microarray spotter for testing stem cell fates inside various miniaturized cell-laden gels in a systematic manner. To demonstrate the feasibility of our platform, we evaluated the osteogenic differentiation of human mesenchymal stem cells (hMSCs) within combinatorial 3D niches. We were able to identify specific combinations, that enhanced the expression of osteogenic markers. Notably, these 'hit' combinations directed hMSCs to form mineralized tissue when conditions were translated to 3D macroscale hydrogels, indicating that the miniaturization of the experimental system did not alter stem cell fate. Overall, our findings confirmed that the 3D cell-laden gel microarray can be used for screening of different conditions in a rapid, cost-effective, and multiplexed manner for a broad range of tissue engineering applications.
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA [2] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [3] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [4] Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark.
Alireza
A
Dolatshahi-Pirouz
1] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [2] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Mehdi
M
Nikkhah
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA [2] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [3] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [4] David H. Koch Institute of Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [5].
Akhilesh K
AK
Gaharwar
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA [2] School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA [3] Vascular Biology Program, Departments of Pathology and Surgery, Children's Hospital Boston, Harvard Medical School, Boston, MA 02115, USA.
Basma
B
Hashmi
1] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [2] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Enrico
E
Guermani
1] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [2] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Hamed
H
Aliabadi
1] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [2] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Gulden
G
Camci-Unal
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
Thomas
T
Ferrante
Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark.
Morten
M
Foss
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA [2] School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA [3] Vascular Biology Program, Departments of Pathology and Surgery, Children's Hospital Boston, Harvard Medical School, Boston, MA 02115, USA.
Donald E
DE
Ingber
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA [2] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02139, USA [3] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Ali
A
Khademhosseini
D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D010012:Osteogenesis / Q000502:physiology*; D046888:Tissue Array Analysis / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods
D020100:Hydrogels