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Device and method for inducing pluripotent cells using energy

A pluripotent cell and energy technology, applied in artificially induced pluripotent cells, non-embryonic pluripotent stem cells, tissue cell/virus culture devices, etc.

Active Publication Date: 2021-07-27
金旼贞 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The exact molecular mechanisms to approximate these approaches are not yet known, so these approaches can only be accepted as alternatives that can achieve safety without the need for the introduction of genetic material, chemicals, and small molecules

Method used

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  • Device and method for inducing pluripotent cells using energy
  • Device and method for inducing pluripotent cells using energy
  • Device and method for inducing pluripotent cells using energy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Preparation of Physics Cells

[0119] figure 1 Is a Physics of the present invention p Luripotent SP h ere y Ielded by Ultra s on IC s Timulus) Cell formation simulation map, with 5W / cm 2 Intensity ultrasonic treatment for 10 minutes of ES (Embryonic STEM) medium mixed human skin fibroblast (HDFA, Cat.No.c-013-5c, Gibco (Invitrogen Cellculture) (1 × 10 6 ), Use 1W / cm 2The intensity ultrasonic treatment of a mixture containing cells for 5 seconds. After screening the survived cells, in the 35 mm bacteria in Petri Dish, the cultured culture was suspended in human ES medium 2 × 10 5 HDF for 6 days.

[0120] The spheroid was formed on the first day of the culture, and the label expression was not differentiated after 3 days.

experiment example 1

[0121] Best Condition Test of Spherical Body Formation

[0122] Human skin fibroblasts form a spheroid in ultrasonic treatment, in order to determine optimal conditions for improving the formation efficiency of the spheroid, alter the ultrasonic treatment conditions, cell culture mode, and the like.

[0123] The cell culture method is used in the surface without a coated cell culture dish culture, a suspension culture medium in the petri dish, and a cell culture which is more easily adhered to the surface on the surface. Monolayer Culture, cultured in dishes (tissue culture dishes).

[0124] Furthermore, the control component is a control group that does not perform any treated control group (NULL) for ultrasonic treatment (UM: Ultrasound Treated Media, 5W / cm) 2 The intensity ultrasonic treatment is 10 minutes of ultrasonic treatment of cells (UC: Ultrasound Treated Cell, 1W / cm) 2 The intensity ultrasonic treatment for 5 seconds) and the cellular and medium of the cell and me...

Embodiment 2

[0163] PHysics Cell Proliferation and Multi - Division

[0164] PHYSICS cell proliferation capacity was evaluated by proliferating marker protein Ki-67 immunostaining and time differential cell nuclear staining with tubicaic acid (HOECHST 33342) and propidium (Pi).

[0165] like Figure 19 As shown, the expression of Ki-67 in Physics cells was detected on day 5.

[0166] In order to clearly prove the proliferation capabilities of PHYSICS cells, the cell proliferation was demonstrated by the following method. Hoechst 33342 with a nuclear nucleus of permeability dyeable survival cells was stained to cultivate Physics cells in the 5th day, completely removed the stained reagent, cultured for 3 days, and cultured 8 days of PHysics cells were fixed with 4% polymethylene formaldehyde. The nucleus is dyed by using PI. The non-repetitive red signal means that new Physics cells were formed due to cell division after 5 days. Further, a single spheroid was cultured, and the pHYSICs were prov...

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Abstract

The present invention relates to a device and method for inducing pluripotent cells using energy. More specifically, energy such as ultrasound, laser or heat treatment is applied to differentiated cells to induce new pluripotent cells with pluripotency.

Description

Technical field [0001] The present invention relates to multi-energy cell inducing devices and methods utilizing energy, by providing ultrasonic waves, laser or heat treatment, etc., can induce multivalent cells. Background technique [0002] Multi-Volume (PLURPOTENCY) is a differentiation into a 3-embryonic system, that is, the ability of foreign embryo, mesocily embryo and endostatic leaves. Multivariate stem cells are important in clinical disease models and transplants because they form any cell or tissue type in the body. Therefore, the main demand in the reprogramming or differentiation of embryonic PLURPOTENT STEM CELL, IPSCs, somatic cells and patients, for clinical applications, no need to introduce foreign genetic substances or Chemicals or small molecules require simple, fast, effective and safe. In recent studies, the interaction between the environment and genotypes is proved to be closely related to gene expression and phenotypic variation of the living body. Cell d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074C12N13/00C12M1/42C12M1/36C12M3/00
CPCC12M41/30C12N5/0696C12N13/00C12N2529/10C12N2523/00C12N2527/00C12N2506/30C12N2506/1307C12M1/42C12M3/00C12M41/14C12M31/00C12M41/46C12N2521/10C12N2501/115C12N2500/32C12N2500/44
Inventor 金舜学
Owner 金旼贞