Construction method and application of research model of bystander effector damaged hematopoietic stem progenitor cells
A bystander effect and research model technology, applied in the field of research model construction, can solve the problems of long-term hematopoietic reconstruction ability, self-renewal ability and decline of hematopoietic progenitor cell clone formation ability in vitro
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Embodiment 1
[0136] In this example, an in vivo research model of bystander effect damage to hematopoietic stem and progenitor cells and an in vivo research model of bystander effect damage to hematopoietic stem and progenitor cells intervened by antioxidant drugs were constructed. The steps are as follows:
[0137] (1) Female immunodeficiency mice (NOG mice) aged 6 to 8 weeks and weighing 18 to 22 g were randomly divided into NR group (control group), IR group (X-ray irradiation group), IR+NAC group group (pretreatment with NAC before X-ray irradiation), IR+SF group (pretreatment with Sulforaphane before X-ray irradiation), and IR+Res group (pretreatment with Resveratrol before X-ray irradiation);
[0138] (2) Mice in IR+NAC group, IR+SF group and IR+Res group were treated with antioxidant drugs 7 days before irradiation, and the treatment plan was as follows:
[0139] IR+NAC group: intraperitoneal injection, 100mg / kg / day, once a day, for 7 consecutive days, and at the same time fed water...
Embodiment 2
[0157] In this example, an in vitro research model of bystander effect damage to hematopoietic stem and progenitor cells and an in vitro research model of bystander effect damage to hematopoietic stem and progenitor cells intervened by antioxidant drugs were constructed. The steps are as follows:
[0158] (1) Configure complete medium for stem cells (fetal bovine serum containing 10% volume fraction, 50ng / mL TPO, 50ng / mL SCF and 50ng / mL Flt-3, the balance being IMDM medium), and containing NAC (100μM), Stem cell complete medium of SF (2.5 μM) and Res (0.1 μM);
[0159] (2) Isolate human bone marrow cells, lyse the red blood cells, and divide them into NR group (control group), IR group (X-ray irradiation group), IR+NAC group (NAC pretreatment before X-ray irradiation), IR+SF group (pretreatment with Sulforaphane before X-ray irradiation) and IR+Res group (pretreatment with Resveratrol before X-ray irradiation), 30 minutes before irradiation, bone marrow cells were resuspended ...
Embodiment 3
[0165] In this embodiment, the human CD34 of the IR group and the NR group in the embodiment 1 subjected to RIBE + The cells were transplanted into mice, and the implantation and differentiation of the cells were detected. The steps are as follows:
[0166] (1) Female NOG mice aged 6 to 8 weeks and weighing 18 to 22 g were randomly divided into groups, and the mice were irradiated with 2Gy of X-rays at a dose rate of 1.2Gy / min the day before transplantation;
[0167] (2) The human CD34 of each group obtained by sorting + Cells were transplanted into mice irradiated with 2Gy through the tail vein, and human CD45 in peripheral blood of mice was continuously detected 4 to 20 weeks after transplantation. + cell ratio;
[0168] (3) 20 weeks after transplantation, the mice were killed by decapitation, and the bone marrow cells of the mice were collected 1×10 6 After washing with staining buffer, resuspend, add antibody, incubate at 4°C in the dark for 30 min, wash once with 2 mL ...
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