US 12,441,969 B2
Delivery of materials to anucleate cells
Armon R. Sharei, Cambridge, MA (US); Klavs F. Jensen, Lexington, MA (US); James Robbins Abshire, Cambridge, MA (US); and Jacquin Clarence Niles, Arlington, MA (US)
Assigned to Massachusetts Institute of Technology, Cambridge, MA (US)
Filed by Massachusetts Institute of Technology, Cambridge, MA (US)
Filed on Mar. 8, 2022, as Appl. No. 17/689,745.
Application 17/689,745 is a division of application No. 15/865,901, filed on Jan. 9, 2018, granted, now 11,299,698.
Application 15/865,901 is a continuation of application No. PCT/US2016/041653, filed on Jul. 8, 2016.
Claims priority of provisional application 62/190,677, filed on Jul. 9, 2015.
Prior Publication US 2022/0195364 A1, Jun. 23, 2022
Int. Cl. C12N 15/87 (2006.01); A61K 35/18 (2015.01); C12M 1/42 (2006.01); C12M 3/06 (2006.01); C12N 5/078 (2010.01)
CPC C12M 23/16 (2013.01) [A61K 35/18 (2013.01); C12M 35/04 (2013.01); C12N 5/0641 (2013.01); C12N 5/0644 (2013.01); C12N 15/87 (2013.01); C12N 2521/00 (2013.01); C12N 2527/00 (2013.01)] 20 Claims
 
1. A composition comprising a mammalian anucleate cell comprising an intracellular payload, wherein the composition is prepared by a method comprising passing a suspension comprising the anucleate cell through a microfluidic channel that includes a cell-deforming constriction causing perturbations of a membrane of the anucleate cell large enough for the payload to pass through, wherein the constriction comprises a width, a length, and a depth, wherein the width of the constriction is less than 4 micrometers; and incubating the anucleate cell in a solution comprising the payload for a predetermined time before or after the anucleate cell passes through the constriction, wherein the anucleate cell is one or more of red blood cells and reticulocytes.