US 12,070,746 B2
Method of osteogenic differentiation in microfluidic tissue culture systems
Dan Gazit, Los Angeles, CA (US); Gadi Pelled, Los Angeles, CA (US); Zulma Gazit, Los Angeles, CA (US); Dmitriy Sheyn, Los Angeles, CA (US); Christopher David Hinojosa, Cambridge, MA (US); Norman Wen, West Roxbury, MA (US); and Geraldine Hamilton, Boston, MA (US)
Assigned to EMULATE, Inc., Boston, MA (US); and CEDARS-SINAI MEDICAL CENTER, Los Angeles, CA (US)
Filed by EMULATE,INC., Boston, MA (US); and CEDARS-SINAI MEDICAL CENTER, Los Angeles, CA (US)
Filed on Apr. 20, 2018, as Appl. No. 15/958,142.
Application 15/958,142 is a continuation in part of application No. PCT/US2017/013250, filed on Jan. 12, 2017.
Claims priority of provisional application 62/277,857, filed on Jan. 12, 2016.
Prior Publication US 2018/0305668 A1, Oct. 25, 2018
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/077 (2010.01); A61K 49/00 (2006.01); A61K 51/02 (2006.01); A61L 27/38 (2006.01); B01L 3/00 (2006.01); C12N 5/00 (2006.01); C12N 5/071 (2010.01); C12N 5/074 (2010.01); C12N 5/0775 (2010.01); G01N 33/50 (2006.01); G01T 1/164 (2006.01); G01T 1/29 (2006.01); C07K 14/78 (2006.01); C12M 1/34 (2006.01)
CPC B01L 3/5027 (2013.01) [A61K 49/0019 (2013.01); A61K 49/005 (2013.01); A61K 51/02 (2013.01); A61L 27/3895 (2013.01); B01L 3/5085 (2013.01); C12N 5/0062 (2013.01); C12N 5/0075 (2013.01); C12N 5/0654 (2013.01); C12N 5/0655 (2013.01); C12N 5/0662 (2013.01); C12N 5/0663 (2013.01); C12N 5/0668 (2013.01); C12N 5/0696 (2013.01); C12N 5/0697 (2013.01); G01N 33/5038 (2013.01); G01T 1/1644 (2013.01); G01T 1/2985 (2013.01); A61M 2205/3334 (2013.01); C07K 14/78 (2013.01); C12M 41/40 (2013.01)] 18 Claims
 
1. A method of osteogenic differentiation comprising
a) seeding mesenchymal stem cells (MSCs) in a microfluidic device in growth medium containing doxycycline (DOX) in the absence of flow to produce attached MSCs,
b) flowing the growth medium in the absence of Bone Morphogenetic Protein-2 (BMP2) such that the attached MSCs proliferate to produce proliferated MSCs,
c) contacting the proliferated MSCs with an osteogenic medium containing Bone Morphogenetic Protein-2 (BMP2) to produce differentiated cells that express one or more of osteocalcin, bone sialoprotein (bsp), osteopontin (opn), and collagen type 1, and
d) providing constant flow of said osteogenic medium during the contacting step to produce a higher level of expression of at least one of said osteocalcin, said bone sialoprotein (bsp), said osteopontin (opn), and said collagen type 1 in said differentiated cells in the presence of said flowing of the osteogenic medium compared to in the absence of said flowing of the osteogenic medium.