Method and compositions for improving allogeneic hematopoietic cell transplantation
A hematopoietic system and cell technology, which is applied in the pharmaceutical application field of homologous hematopoietic system reconstituted cells
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Embodiment 1
[0061] Example 1. Transplant Patient Checkup
[0062] The investigation of allografts or allografts was characterized in that it was performed on 104 patients who had received non-T cell depleted allogeneic bone marrow transplants from HLA-matched siblings. Pretransplant status adjustments for these patients included butyl dimesylate treatment (n=89), total body irradiation with administration of cyclophosphamide (n=10), and cyclophosphamide (n=5). The total number of cells in the graft averaged 3 × 10 8 Cells / kg recipient body weight. CD34 + The number of cells correlated positively with an increase in survival (p=0.0004, overall survival; p=0.001, disease-free survival). The graft was additionally quantified for a cell population characterized by CD3 - , CD4 +(Hi) , CD8 - , low side scatter, here considered DCP, the number of these cells in transplanted species was inversely correlated with overall survival (OS) and disease-free survival (DFS, p=0.005). In this surv...
Embodiment 2
[0064] Example 2. Transplantation of selected stem cells
[0065] donor source : Bone marrow or peripheral blood stem cells collected from HLA-matched or mismatched donors.
[0066] Donor PBSC collection : To immobilize PBSC, G-CSF was administered daily until complete depletion of leukocytes. Donors received subcutaneous injections of G-CSF at 10 μg / kg / day according to their actual body weight. Institutional standards of donor support care must be followed during mobilization and leukapheresis. PBSC were collected by leukapheresis. Leukocyte depletion should be performed on a continuous flow cytometry instrument according to institutional standard methods. The endpoint of each leukapheresis collection should be the processing of more than 12 liters of whole blood or 4 hours of leukapheresis. Leukapheresis should be done after donor G-CSF treatment + Start on day 5 and proceed daily (leukapheresis up to four days) until 2 x 10 6 CD34 + Target cells / kg recipient bo...
Embodiment 3
[0070] Example 3. cell sorting
[0071] General protocol for cell sorting. Cells were stained with FITC-conjugated CD34-specific monoclonal antibodies and PE-conjugated monoclonal antibodies recognizing CD4, CD123, CD36 or CD45RA (Becton-Dickinson, San Jose, CA). A directly conjugated isotype-matched irrelevant antibody was used as a reference. Cells were stained for 30 min at 4°C in a staining buffer consisting of Hank's saline buffered solution (HBSS, Mediatech, Herndon, VA) containing 2% v / v autologous human plasma or 1% human serum albumin. Excess antibody was removed by dilution with 10 volumes of staining buffer followed by centrifugation at 500 xg. Stained cells were analyzed indirectly using a FACSCAN flow cytometer (BDIS, San Jose, CA) or sorted aseptically using a FACSVANTAGE cell sorter (BDIS, San Jose, CA). Central and marginal scatter gates for viable nucleated cells were used to obtain list-mode data. Using CellQuest TM Or LYSIS-II software (BDIS, San Jos...
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