US 2013021 6506A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2013/0216506 A1 DISCHER et al. (43) Pub. Date: Aug. 22, 2013 (54) BIOREACTOR FOR ISOLATION OF RARE A6135/14 (2006.01) CELLS AND METHODS OF USE CI2O I/68 (2006.01) CI2N5/078 (2006.01) (71) Applicant: THE TRUSTEES OF THE CI2N5/0789 (2006.01) UNIVERSITY OF PENNSYLVANIA, GOIN L/10 (2006.01) (US) GOIN33/569 (2006.01) (52) U.S. Cl. (72) Inventors: DENNIS E. DISCHER, CPC ................. CI2M 47/04 (2013.01); G0IN I/10 PHILADELPHIA, PA (US); JAE-WON (2013.01); A61K 35/28 (2013.01); G0IN SHIN, PHILADELPHIA, PA (US) 33/56972 (2013.01); C12O I/6881 (2013.01); (73) Assignee: THE TRUSTEES OF THE CI2N5/0644Sets (2013.O1): SK st CI2N5/0647 (2013.01) UNIVERSITY OF PENNSYLVANIA, 1. PHILADELPHIA PA (US USPC ... 424/93.7:435/309.1:435/304. 1; 435/325; , PA (US) 435/2; 435/7.92; 435/6.12:506/9; 435/34 (21) Appl. No.: 13/772,750 (22) Filed: Feb. 21, 2013 (57) ABSTRACT Related U.S. Application Data The present invention relates to a bioreactor apparatus, and (60) Provisional application No. 61/601,364, filed on Feb. methods of use, for the isolation of rare blood cells, including 21, 2012. hematopoietic stem cells and megakaryocytes. The apparatus includes a Soft Substrate and an anti-contractility agent, Publication Classification thereby providing a soft microenvironment to cultured cells. The apparatus of the invention is permissive for the survival (51) Int. Cl. of non-dividing cells while dividing cells are eliminated. This CI2M I/00 (2006.01) unique property allows for the simple isolation of rare blood A6 IK35/28 (2006.01) cells without the use of costly equipment and antibodies. 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Patent Application Publication Aug. 22, 2013 Sheet 55 of 55 US 2013/021650.6 A1 ?ooogººpoole-o! 99eun61– ***********?,¿ &&&&&&&&&&&&&&&&&&&&&««·*…***-3· &*******??&&&??ae,§§ US 2013/021650.6 A1 Aug. 22, 2013 BOREACTOR FOR SOLATION OF RARE methods that isolate, use and assay rare blood cells. The CELLS AND METHODS OF USE present invention addresses these unmet needs in the art.