Cell reprogramming method using physical stimulation-mediated environmental influx
The present invention relates to a cell reprogramming method using a physical stimulation-mediated environmental influx, and more specifically, by subjecting differentiated or non-differentiated cells to physical stimulation which can promote an environmental influx, such as ultrasonic waves, laser or heat shock, without the introduction of a reprogramming-inducing factor or a chemical substance to the differentiated cells, the cells can be reprogrammed with just the imposition of an external environmental influx into pluripotent cells or arbitrary differentiated cells having a different expression type from the differentiated or non-differentiated cells, and as such an inducement has a simple and effective production process, the possibility of an autogenic cell therapy can be made greater.
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/cm 2 to 3 W/cm 2 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.
2. The cell reprogramming method of claim 1 , wherein the environmental influx includes an intracellular influx of genetic materials, chemicals, small molecules, exosomes, or extracellular vesicles containing exosomes released from differentiated or non-differentiated cells subjected to the ultrasonic waves treatment; or culture medium components.
3. The cell reprogramming method of claim 1 , further comprising:
subjecting the culture medium to the ultrasonic waves treatment having an output intensity of 1 W/cm 2 to 20 W/cm 2 for 1 to 20 minutes before mixing the HDF with the culture medium.
4. The cell reprogramming method of claim 1 , wherein the mixture subjected to the ultrasonic waves treatment is cultured for 1 to 20 days by a suspended culture or monolayer culture method.