[methods and apparatuses for isolating and preparing stem cells]

Abstract

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.