A Size-Selective Intracellular Delivery Platform.
Identifying and separating a subpopulation of cells from a heterogeneous mixture are essential elements of biological research. Current approaches require detailed knowledge of unique cell surface properties of the target cell population. A method is described that exploits size differences of cells to facilitate selective intracellular delivery using a high throughput microfluidic device. Cells traversing a constriction within this device undergo a transient disruption of the cell membrane that allows for cytoplasmic delivery of cargo. Unique constriction widths allow for optimization of delivery to cells of different sizes. For example, a 4 μm wide constriction is effective for delivery of cargo to primary human T-cells that have an average diameter of 6.7 μm. In contrast, a 6 or 7 μm wide constriction is best for large pancreatic cancer cell lines BxPc3 (10.8 μm) and PANC-1 (12.3 μm). These small differences in cell diameter are sufficient to allow for selective delivery of cargo to pancreatic cancer cells within a heterogeneous mixture containing T-cells. The application of this approach is demonstrated by selectively delivering dextran-conjugated fluorophores to circulating tumor cells in patient blood allowing for their subsequent isolation and genomic characterization.
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.|Hospital Medicine Unit, Department of Medicine, Massachusetts General Hospital, Boston, MA, 02114, USA.
May Tun
MT
Saung
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Armon
A
Sharei
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Viktor A
VA
Adalsteinsson
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Nahyun
N
Cho
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Tushar
T
Kamath
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Camilo
C
Ruiz
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Jesse
J
Kirkpatrick
Advanced Tissue Resources Core, Massachusetts General Hospital, Charlestown Navy Yard, Charlestown, MA, 02129, USA.
Nehal
N
Patel
Department of Pathology, Massachusetts General Hospital, Boston, MA, 02114, USA.
Mari
M
Mino-Kenudson
Department of Surgery, Massachusetts General Hospital, Boston, MA, 02114, USA.
Sarah P
SP
Thayer
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Robert
R
Langer
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.
Klavs F
KF
Jensen
Department of Surgery, Massachusetts General Hospital, Boston, MA, 02114, USA.
Andrew S
AS
Liss
Department of Chemical Engineering, Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, 02139, USA.|The Broad Institute of Harvard and MIT, Cambridge, MA, 02142, USA.
J Christopher
JC
Love
circulating tumor cells; intracellular delivery; microfluidics; size-selective delivery