Methods and apparatuses for isolating and preparing stem cells
The present invention relates to a method of preparing a population of stem cells for autologous implantation to a subject. 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. Also the invention relates to therapeutic applications of the activated stem cells.
1. A method of preparing a population of adipose-derived stem cells (ADSC) for autologous implantation to a subject, comprising:
(i) collecting adipose tissue and blood from a subject,
(ii) isolating ADSCs cells from the adipose tissue and platelet rich plasma (PRP) from the blood,
(iii) activating the isolated ADSCs by irradiation with yellow light at a wavelength of 575-595 nm and at 5-20 mW, red light at wavelength of 630-635 nm and at 10-100 mW, and green light at a wavelength of 510-540 nm and at 10-60 mW, for 30-60 minutes,
(iv) mixing the activated ADSCs with the PRP, thereby preparing a population of ADSCs for autologous implantation.
2. The method of claim 1 , wherein the ADSCs are irradiated with yellow light at a wavelength of 595 nm and at 20 mW, red light at wavelength of 635 nm and at 60 mW, and green light at a wavelength of 535 nm and at 60 mW, for 30-60 minutes.
3. The method of claim 1 , wherein the isolated ADSCs in step (ii) are exposed to one or more growth factors.
4. The method of claim 3 wherein the growth factors are Epidermal Growth Factor (EGF); Platelet-Derived Growth Factor (PDGF); Fibroblast Growth Factor (FGF); Transforming Growth Factor-β (TGF-β); Erythropoietin (EPO); Insulin-like Growth Factor-I (IGF-I); Insulin-like Growth Factor-II; and/or Tumor Necrosis Factor-α (TNF-α).
5. The method of claim 3 wherein the PRP comprises one or more growth factors.
6. The method of claim 1 , wherein step (i) comprises exposing the adipose tissue to a lipid dissolving agent.
7. The method of claim 6 wherein said lipid dissolving agent is lecithin.
8. The method of claim 1 , wherein step (i) comprises exposing the adipose tissue to collagenase.
9. The method of claim 1 , wherein the adipose tissue is isolated from the subject by liposuction and/or lipoplasty.
10. The method of claim 1 comprising:
i) processing the 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; and
ii) activating the population of ADSC by irradiating the cells with yellow light at a wavelength of 575-595 nm and at 5-20 mW, red light at wavelength of 630-635 nm and at 10-100 mW, and green light at a wavelength of 510-540 nm and at 10-60 mW, for 30-60 mins, and incubating the ADSCs in the presence of PRP prepared from the patient to be treated at 30° C. to 38° C. for 5-120 minutes.