[METHODS AND APPARATUSES FOR ISOLATING AND PREPARING STEM CELLS]
A method of preparing a population of stem cells for autologous implantation to a subject and apparatus used therein are provided. The cells are activated by irradiating the cells with one or more wavelengths of yellow and red and/or green light. In particular, the cells are irradiated with 575-595 nm (5-20 mW), and 630-635 nm or 660-670 nm (10-100 mW) and/or 510-540 nm (10-60 mW) of monochromatic light for 30-60 mins. Preferably the stem cells are adipose-derived stem cells. Therapeutic applications of the activated stem cells are also provided.
1 - 35 . (canceled)
36 . A method of preparing a population of stem cells for autologous implantation to a subject, comprising activating the stem cells by irradiating the cells with one or more wavelengths of yellow and red and/or green light.
37 . The method of claim 36 , wherein yellow and red and green light is used.
38 . The method of claim 36 , wherein the cells are irradiated with 575-595 nm (5-20 mW), and 630-635 nm (10-100 mW) and/or 510-540 nm (10-60 mW) of monochromatic light for 30-60 mins.
39 . The method of claim 38 , wherein the cells are irradiated with 595 nm (20 mW), 635 nm (60 mW) and 535 nm (60 mW) of monochromatic light, for 30-60 mins.
40 . The method of claim 36 , comprising a further initial step of processing a sample of tissue from the subject to obtain the population stem cells to be irradiated.
41 . The method of claim 36 , wherein the stem cells are adult stem cells.
42 . The method of claim 41 , wherein the adult stem cells are adipose-derived stem cells (ADSC); dermal stem cells; hematopoietic stem cells; mammary stem cells; mesenchymal stem cells; endothelial stem cells; neural stem cells; neural crest stem cells; or testicular stem cells.
43 . The method of claim 36 , wherein the stem cells are ADSC and the tissue is adipose tissue.
44 . The method of claim 36 , wherein the stem cells are exposed to one or more growth factors.
45 . The method of claim 44 , wherein the growth factors are Epidermal Growth Factor (EGF); Platelet-Derived Growth Factor (PDGF); Fibroblast Growth Factor (FGFs); Transforming Growth Factors-b (TGFs-b); Erythropoietin (EPO); Insulin-like Growth Factor-I (IGF-I); Insulin-like Growth Factor-II; and/or Tumour Necrosis Factor-a (TNF-a).
46 . The method of claim 44 , wherein the growth factors are provided by platelet-rich plasma prepared from the subject.
47 . The method of claim 43 , wherein the step of processing the sample of adipose tissue includes exposing said tissue to a lipid dissolving agent.
48 . The method of claim 47 , wherein said lipid dissolving agent is lecithin.
49 . The method of claim 43 , wherein the step of processing the sample of adipose tissue includes exposing said tissue to collagenase.
50 . The method of claim 40 , wherein where the tissue is adipose tissue, then the adipose tissue has been isolated from the subject by liposuction and/or lipoplasty.
51 . The method of claim 43 , comprising:
i) processing a sample of adipose tissue from the subject to obtain a concentrated population of ADSC, said processing comprising: removing free lipid and single cell components of the tissue by rinsing; disaggregating the tissue using mechanical forces, collagenase and lecithin digestion; separating and concentrating the ADSC using centrifugation;
ii) activating the population of ADSC by irradiating the cells with 575-595 nm (5-20 mW), and 630-635 nm (10-100 mW) and/or 510-540 nm (10-60 mW) of monochromatic light for 30-60 mins, and incubating the cells in the presence of platelet-rich plasma prepared from the patient to be treated at 30° C. to 38° C. for 5-120 mins.
52 . The method of claim 36 , wherein said method does not involve a step of culturing the population of stem cells to increase cell number prior to their use in autologous implantation.
53 . A population of stem cells for autologous implantation obtained by the method of claim 36 .
54 . A method of treating a patient using stem cells for autologous implantation comprising:
(i) isolating tissue from the patient;
(ii) preparing a population of stem cells according to a method of claim 36 ; and
(iii) administering the activated population of stem cells to the patient.
55 . The method of claim 54 , wherein the population of stem cells is mixed with a further portion of tissue prior to administration to the patient.
56 . The method of claim 54 , wherein the stem cells are ADSC and the tissue is adipose tissue.
57 . The method of claim 54 , wherein said patient is treated for therapeutic and/or cosmetic purposes.
58 . A population of stem cells prepared according to the method of claim 36 for therapeutic and/or cosmetic purposes.
59 . A population of stem cells prepared according to the method of claim 54 for therapeutic and/or cosmetic purposes.
60 . The method of claim 54 , wherein said treatment is for bone-related disorders; adipose related disorders or diseases; liver related diseases; myocardial infarctions; renal diseases; retinal diseases; wound healing; skeletal muscle disorders; cartilage and joint repair; lung injuries; diabetes; intestinal disorders; nervous system disorders, diseases, or injuries; diabetes; alopecia.
61 . The method of claim 60 , wherein the treatment is for type II diabetes.
62 . A method of treating a subject comprising activating stem cells in situ in the subject by irradiating the cells with one or more wavelengths of yellow and red and/or green light.
63 . The method of claim 62 , wherein the cells are irradiated in situ in the subject with 575-595 nm (5-20 mW), and 630-635 nm (10-100 mW) and/or 510-540 nm (10-60 mW) of monochromatic light for 30-60 mins.
64 . The method of claim 63 , wherein the cells are irradiated in situ in the subject with 595 nm (20 mW), 635 nm (60 mW) and 535 nm (60 mW) of monochromatic light, for 30-60 mins.
65 . An apparatus for the preparation of a population of stem cells for autologous implantation, said apparatus comprising:
i) a tissue preparation container including a one way inlet port structured to receive tissue removed from a patient and for the administration of at least one additive to mix with the stem cells contained therein;
ii) an outlet structured to permit the cells in the container to be removed.
66 . The apparatus of claim 65 , further comprising means for mechanically disrupting the tissue.
67 . The apparatus of claim 65 , further comprising means for separating, and optionally concentrating, the stem cells from further components of the tissue.
68 . The apparatus of claim 65 , further comprising
i) one or more sources of wavelengths of yellow and red and/or green light; and,
ii) a locator for arranging a population of cells in the said wavelengths of light.
69 . An apparatus for preparing an activated population of stem cells for autologous implantation to a subject, comprising:
i) one or more sources of wavelengths of yellow and red and/or green light; and,
ii) a locator for arranging a population of cells in the said wavelengths of light.
70 . The apparatus of claim 68 , wherein the wavelengths are 575-595 nm (5-20 mW), and 630-635 nm (10-100 mW) and/or 510-540 nm (10-60 mW) of monochromatic light; most preferably 595 nm (20 mW), 635 nm (60 mW) and 535 nm (60 mW) of monochromatic light.
71 . The apparatus of claim 69 , wherein the wavelengths are 575-595 nm (5-20 mW), and 630-635 nm (10-100 mW) and/or 510-540 nm (10-60 mW) of monochromatic light; most preferably 595 nm (20 mW), 635 nm (60 mW) and 535 nm (60 mW) of monochromatic light.
72 . The apparatus of claim 65 , wherein the stem cells are ADSC and the tissue is adipose tissue.
73 . The apparatus of claim 69 , wherein the stem cells are ADSC and the tissue is adipose tissue.