US 11,859,175 B2
Cell reprogramming method using physical stimulation-mediated environmental influx
Soon Hag Kim, Seongnam-si (KR)
Assigned to STEMON INC., Seongnam-si (KR)
Appl. No. 16/082,926
Filed by STEMON INC., Seongnam-si (KR)
PCT Filed Aug. 9, 2016, PCT No. PCT/KR2016/008754
§ 371(c)(1), (2) Date Dec. 26, 2018,
PCT Pub. No. WO2017/155166, PCT Pub. Date Sep. 14, 2017.
Claims priority of application No. 10-2016-0029611 (KR), filed on Mar. 11, 2016; and application No. 10-2016-0071852 (KR), filed on Jun. 9, 2016.
Prior Publication US 2019/0119666 A1, Apr. 25, 2019
Int. Cl. C12N 5/00 (2006.01); C12N 13/00 (2006.01); C12N 5/0797 (2010.01); C12N 5/077 (2010.01); C12N 5/071 (2010.01)
CPC C12N 13/00 (2013.01) [C12N 5/067 (2013.01); C12N 5/0623 (2013.01); C12N 5/0656 (2013.01); C12N 5/0658 (2013.01); C12N 2521/10 (2013.01); C12N 2523/00 (2013.01); C12N 2529/00 (2013.01)] 4 Claims
 
1. A cell reprogramming method comprising:
subjecting a mixture of human dermal fibroblasts (HDF) and a culture medium to ultrasonic waves treatment having an output intensity of 0.5 W/cm2 to 3 W/cm2 for 1 to 5 seconds which promote an environmental influx for a cell reprogramming, wherein the ultrasonic waves treatment induces damage of the cell membrane which is recovered 2 hours after being subjected to the ultrasonic waves treatment; and
culturing, immediately after the ultrasonic waves treatment, the mixture subjected to the ultrasonic waves treatment for a predetermined time to obtain reprogrammed cells,
wherein the culture medium is any one of a hepatocyte differentiation-inducing medium, an adipocyte differentiation-inducing medium, or a neuronal cell differentiation-inducing medium,
wherein the ultrasonic treatment promotes the environmental influx without introduction of a reprogramming inducing factor other than any one of the differentiation-inducing medium,
wherein the reprogrammed cells are any one of neurons expressing any one of PAX6, Nestin, Sox1, Sox2, MAP2, TuJl, GFAP or O4; hepatocytes expressing any one of AFP, HNF1a, HNF4a, CK18 or ALB; or adipocytes stained with oil red O and expressing any one of Pparc2, C/ebpa, aP2 or Fabp4.