A new and useful method is provided, for enhancing the immune system of a donor/recipient. Stem cells are collected from the donor/recipient at a time that the donor/recipient appears to have a healthy immune system, taking into consideration the age of the donor/recipient and the characteristics of a healthy immune system for a donor/recipient of that age. The stem cells are processed and stored in a condition that enables the stem cells to be later administered to that donor/recipient in predetermined amounts, in serial (consecutive) fashion, over predetermined time intervals, and at the later request of the donor/recipient, the stem cells are administered to the donor/recipient in serial (consecutive) fashion, over predetermined time intervals, to enhance the donor/recipient's immune system thereby preventing formation of age related diseases that are caused by rundown immune system. The practice of the present invention results in replenishing the hematopoietic system and primarily rebuilding the immune system in an aged individual. Also noteworthy, it refreshes the repertoire of stem cells, which are at the point of exhaustion, bringing about extension of life span and can also be used for improvement of cognitive functions of the donor/recipient.
F.02 Acquisition of new scientific knowledge
COBISS.SI-ID: 33446105The molecular mechanisms involved in aging of stem cells are the same as ones involved in aging of somatic cells, including telomere shortening, oxidative stress, epigenetic dysregulation, miRNAs changes, and altered proteome. Various pathways, such as insulin/IGF-1, mTOR, FoxO, AMP-activated protein kinase (AMPK), sirtuin, and many others are associated with aging. Senescent stem cells exhibit functional and numerical impairment, including decreased responsiveness to physical environment, decreased resistance to oxidative stress and deterioration of the pluripotent stem cell pool. The most evident consequences are found in the immune system, where both innate as well as the adaptive immunity are affected, exhibiting a plethora of defects of macrophages, dendritic cells, NK-cells, granulocytes, as well as in B and T lymphocytes. The final result is defective immune response with autoimmune tensions, increased tolerance to cancer and chronic oxi-inflammatory status. For the regenerative purposes, stem cells must better be devoid of senescent defects. Stem cell therapies mandate the best quality of stem cells grafts. With this respect, biology of senescence of stem cells has to be taken into account in every stem cell based therapy, as well as in tissue engineering procedures.
F.02 Acquisition of new scientific knowledge
COBISS.SI-ID: 33749721Rationale: Preclinical data in heart failure models suggest that repetitive stem cell therapy may be superior to single-dose cell administration. Objective: We investigated whether repetitive administration of CD34+ cells is superior to single dose administration in patients with non-ischemic dilated cardiomyopathy (DCM). Methods and Results: Of 66 patients with DCM, NYHA functional class III, and left ventricular ejection fraction (LVEF)( 40% enrolled in the study, 60 were randomly allocated to repetitive cell therapy (Group A, N=30), or single cell therapy (Group B, N=30). Patients received granulocyte-colony stimulating factor (G-CSF) for 5 days and 80 million CD34+ cells were collected by apheresis and injected transendocardially. In Group A, cell therapy was repeated at 6 months. All patients were followed for 1 year, and the primary end-point was the difference in change in LVEF between the groups. At baseline, the groups did not differ in age, sex, LVEF, NT-proBNP, or 6-minute walk test distance. When directly comparing groups A and B at 1 year, there was no significant difference in change in LVEF (from 32.2±9.3% to 41.2±6.5% in Group A and from 30.0±7.0% to 37.9±5.3% in Group B, P=0.40). From baseline to 6 months, both groups improved in LVEF (+6.9±3.3% in Group A, P=0.001 and +7.1±3.5% in Group B, P=0.001), NT-proBNP (-578±211 pg/ml, P=0.02 and -633±305 pg/ml, P=0.01) and 6MWT (+87±21 m, P=0.03 and +92±25 m, P=0.02). In contrast, we observed no significant changes between 6 months and 1 year (LVEF: +2.1±2.3% in Group A, P=0.19 and +0.8±3.1% in Group B, P=0.56; NT-proBNP: -215±125 pg/ml, P=0.26 and -33±205 pg/ml, P=0.77; 6MWT: +27±11 m, P=0.2 and +12±18 m, P=0.42). Conclusions: In patients with DCM, repetitive CD34+ cell administration does not appear to be associated with superior improvements in LVEF, NT-proBNP, or 6MWT when compared to single dose cell therapy.
F.21 Development of new health/diagnostic methods/procedures
COBISS.SI-ID: 5052844Mesenchymal stem cells (MSCs) are heterogeneous population of cells with great potential for regenerative medicine. MSCs are relatively easy to expand in a cell culture, however determination of their concentration in harvested tissue is more complex and is not implemented as routine procedure. To identify MSCs collected from bone marrow we have used two combinations of cell markers (CD45-/CD73+/CD90+/CD105+ and CD45-/CD271+) and fibroblast colony-forming unit (CFU-F) assay. Further, in donors of various ages, mesenchymal stem cell concentration was compared with the result of CFU-F assay and with hematopoietic stem cell concentration, determined by a standardized flow cytometric assay. A positive correlation of MSC populations to the CFU-F numbers is observed, the population of the CD45-/CD271+ cells correlates better with CFU-F numbers than the population of the CD45-/CD73+/CD90+/CD105+ cells. The relationship between the hematopoietic CD45dim/CD34+ cell concentration and mesenchymal CFU-Fs or CD45-/CD271+ cells shows a positive linear regression. An age-related quantitative reduction of hematopoietic CD45dim/CD34+, mesenchymal CD45-/CD73+/CD90+/CD105+ and CD45-/CD271+ stem cells, and CFU-F numbers were noted. Additionally, statistically significant higher CFU-F numbers were observed when bone marrow samples were harvested from three different sites from the anterior iliac crest instead of harvesting the same sample amount only from one site.
F.21 Development of new health/diagnostic methods/procedures
COBISS.SI-ID: 33951961The aim of the study was to evaluate the efficacy and safety of the new method of platelet-rich plasma activation in the form of platelet gel, used in the treatment of non-healing %chronic lower leg ulcers. The study was prospectively randomized, double blind and placebo controlled. We treated 60 patients (42 males and 18 females, mean age 69.43 years, SD 14.74) with chronic lower leg ulcers of different etiologies. Thirty patients were treated with allogeneic platelet gel and 30 with %hydrogel. Both groups were comparable for duration of ulcer and its size. Treatment was repeated once a week for three consecutive weeks and then the last examination was scheduled at 6 months of the first platelet gel application. The t-test was used to analyze independent samples. Healing of chronic wounds with platelet gel was statistically significantly more effective compared to the treatment with hydrogel (p(0.05). At 6 months of platelet gel application, the mean wound area in the experimental group decreased to 35.01% (SD 53.69) of the initial wound size. In the control group, the wound area decreased to 89.95% (SD 71.82) of the initial wound size (p=0.001). The circumference of the wounds diminished to 54.62% (SD 39.85) of the initial value in the experimental group, compared to 91.28% (SD 29.32) in the control group (p(0.001). Allogeneic platelet gel prepared by the new method used in this study was found to be a good treatment option for non-healing chronic wounds.
F.21 Development of new health/diagnostic methods/procedures
COBISS.SI-ID: 34212057