| Obtaining cellular preparation by isolating lymphocytes, activating in culture flasks or plates with nutrient medium containing L-glutamine and albumin, re-suspending activated cytotoxic lymphocytes, renewing medium and distributing supernatants into ampoules, and storing in freezer | |
| 2024-07-10 | |
| 专利权人 | FEDERAL STATE BUDGETARY INST (FSBI-C) |
| 申请日期 | 2024-07-10 |
| 专利号 | RU2024119288-A; RU2835686-C2 |
| 成果简介 | NOVELTY - Method for obtaining a cellular preparation based on activated lymphocytes from leukocyte filter involves (a) isolating lymphocytes on a Ficoll(RTM: Inert and hydrophilic polysaccharide) gradient with a density of 1.077 g/cm3, (b) activating in culture flasks or plates with a nutrient medium containing recombinant human interleukin-2 (IL-2), interleukin-15 (IL-15), albumin, sodium pyruvate, gentamicin and L-glutamine for 10-14 days in a carbon dioxide incubator, (c) re-suspending activated cytotoxic lymphocytes in the supernatant and performing intradermal paravertebral administration from the 3rd day of activation, and (d) renewing half the volume of the nutrient medium every 48 hours and distributing supernatants into 1.3-2 ml ampoules, and storing in a freezer for subsequent use in outpatient treatment, where donor lymphocytes are obtained from leukocyte filters by backwashing with isotonic sodium chloride solution and a medical drip system. USE - The method is useful for obtaining a cellular preparation based on activated lymphocytes from leukocyte filter. DETAILED DESCRIPTION - Method for obtaining a cellular preparation based on activated lymphocytes from leukocyte filter involves (a) isolating lymphocytes on a Ficoll(RTM: Inert and hydrophilic polysaccharide) gradient with a density of 1.077 g/cm3, (b) activating in culture flasks or plates with a nutrient medium containing recombinant human interleukin-2 (IL-2), interleukin-15 (IL-15), albumin, sodium pyruvate, gentamicin and L-glutamine for 10-14 days in a carbon dioxide incubator, (c) re-suspending activated cytotoxic lymphocytes in the supernatant and performing intradermal paravertebral administration from the 3rd day of activation, and (d) renewing half the volume of the nutrient medium every 48 hours and distributing supernatants into 1.3-2 ml ampoules, and storing in a freezer for subsequent use in outpatient treatment, where donor lymphocytes are obtained from leukocyte filters by backwashing with isotonic sodium chloride solution and a medical drip system, and 180-350 million activated lymphocytes (human leukocyte antigen-DR (HLA-DR)+) are obtained from one filter including T-cytotoxic cells (cluster of differentiation (CD)3+CD8+) and natural killer T (NKT)-cells (CD3+CD16+CD56+) with high viability ≯ 96%. |
| IPC 分类号 | A61K-038/20 ; C12N-005/0781 ; C12N-005/0783 |
| 国家 | 俄罗斯 |
| 专业领域 | 生物科学 |
| 语种 | 英语 |
| 成果类型 | 专利 |
| 文献类型 | 科技成果 |
| 条目标识符 | http://119.78.100.226:8889/handle/3KE4DYBR/15860 |
| 专题 | 中国科学院新疆生态与地理研究所 |
| 作者单位 | FEDERAL STATE BUDGETARY INST (FSBI-C) |
| 推荐引用方式 GB/T 7714 | GRIVTSOVA L YU,GELM YU V,SEMENOVA N S,et al. Obtaining cellular preparation by isolating lymphocytes, activating in culture flasks or plates with nutrient medium containing L-glutamine and albumin, re-suspending activated cytotoxic lymphocytes, renewing medium and distributing supernatants into ampoules, and storing in freezer. RU2024119288-A; RU2835686-C2[P]. 2024. |
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