Method for producing insulin-secreting stem cells and cells derived therefrom
The present invention is a method for producing an insulin-secreting pre-adipocyte stem cell and the cells derived therefrom. The preferred method comprises obtaining fat cells from a human; treating the fat cells with low-level laser energy; culturing the treated fat cells and suspending them in media together with an insulin gene plasmid; and subjecting the fat cells to transfection. The transfection is facilitated with the application of low-level laser energy.
1 . A method for producing an insulin-secreting stem cell, the method comprising: suspending one or more fat cells in media together with an insulin gene and treating the suspended fat cells with laser energy.
2 . The method according to claim 1 wherein the fat cells are from fat tissue or lipoaspirate.
3 . The method according to claim 1 wherein the laser energy is applied for not more than 20 minutes.
4 . The method according to claim 1 wherein the insulin gene is supplied a predetermined concentration.
5 . The method according to claim 4 in which the insulin DNA concentration is 0.5 to 2 micrograms per 1.6 million cells.
6 . The method of claim 1 wherein the insulin gene is carried by a plasmid.
7 . The method according to claim 1 wherein the laser energy is less than 1 watt.
8 . The method according to claim 1 wherein the laser energy is in the visible spectrum.
9 . The method according to claim 1 wherein the laser energy is in the red range of the visible spectrum.
10 . The method according to claim 1 wherein the wavelength of the laser energy is about 635 nm.
11 . The method according to claim 1 wherein the pulse frequency of the laser energy is less than 100,000 Hz.
12 . The method of claim 1 which further comprises:
obtaining the fat cells from a human; and
culturing the fat cells.
13 . The method according to claim 12 wherein the media includes a serum autologous to the human.
14 . The method according to claim 12 wherein culturing further comprises treating the fat cells in culture with laser energy.
15 . The method according to claim 12 wherein the culturing occurs in an incubator.
16 . The method according to claim 15 wherein the incubation is for not more than four weeks.
17 . The method according to claim 12 wherein the laser energy applied to the fat cells in culture is less than 1 watt.
18 . The method according to claim 14 wherein the laser energy applied to the fat cells in culture is in the visible spectrum.
19 . The method according to claim 14 wherein the laser energy applied to the fat cells in culture is in the red range of the visible spectrum.
20 . The method according to claim 13 wherein the laser energy applied to the fat cells in culture is about 635 nm.
21 . The method according to claim 13 wherein the laser energy applied to the fat cells in culture is less than 100,000 Hz.
22 . A method for producing an insulin-secreting pre-adipocyte stem cell, the method comprising:
a. obtaining fat cells from lipoaspirate of a diabetic human;
b. exposing the fat cells with a first laser energy treatment to facilitate the removal of lipid out of the fat cells to create laser-treated cells;
c. culturing the laser-treated cells by washing them with isotonic fluids and placing them in media that includes an serum autologous to the human in an incubator for a defined period of time not exceeding 4 weeks to create cultured cells;
d. suspending the cultured cells in media together with an insulin gene plasmid to create suspended cells;
e. treating the suspended cells with a second low-level laser energy to facilitate the transfection of the insulin gene.
23 . The method according to claim 22 wherein the first and second laser energy treatments use laser sources of less than 1 watt.
24 . The method according to claim 22 wherein the first and second laser energy treatments use laser energy in the visible spectrum.
25 . The method according to claim 22 wherein first and second laser energy treatments use laser energy in the red range of the visible spectrum.
26 . The method according to claim 22 wherein the wavelength of first laser energy treatment is about 635 nm.
27 . The method according to claim 22 wherein the first laser energy treatment uses a pulse frequency of less than 100,000 Hz.
28 . An insulin-secreting pre-adipocyte stem cell.
29 . The cell of claim 28 produced by the method of claim 1 .
30 . The cell of claim 28 produced by the method of claim 22 .
31 . An insulin-secreting stem cell which has been differentiated from stem cells found in fat tissue.
32 . An insulin-secreting stem cell which has been differentiated from stem cells found in lipoaspirate.
33 . A method of treating disease in a human which comprises administering cells derived from the method of claim 1 .
34 . A method of claim 33 in which the human being treated is the same as the human from which the fat cells are obtained.
35 . A method of treating disease in a human which comprises administering cells derived from the method of claim 22 .
36 . A method of claim 35 in which the human being treated is the same as the human from which the fat cells are obtained.
37 . A method of treating disease in a human which comprises administering cells of claim 28 .
38 . A method of treating disease in a human which comprises administering cells of claim 31 .
39 . A method of claim 38 in which the human being treated is the same as the human from which the fat tissue is obtained.
40 . A method of treating disease in a human which comprises administering cells of claim 32 .
41 . A method of claim 40 in which the human being treated is the same as the human from which the lipoaspirate is obtained.