TIL肿瘤浸润淋巴细胞培养基
——LONZA X-VIVO 15 培养基
产品名称：X-VIVO 15 无血清免疫细胞培养基
生产厂家：瑞士LONZA集团
生产地区：美国/比利时
产品规格：1L/瓶
培养细胞：肿瘤浸润淋巴细胞(TIL)、T细胞、CAR-T等
瑞士LONZA集团在细胞免疫治疗基础研究相关产品的研发上卓有建树，旗下X-VIVO 15 培养基，针对TIL肿瘤浸润淋巴细胞在体外培养环境中的增殖进行了优化，大量应用于包括TIL在内的T细胞（T cells）、造血干细胞（Hematopoietic stem cells）、PBL（Peripheral Blood Lymphocytes）、DC（Dendritic cells）、CIK（Cytokine-induced killer）、NK（natural killer cell）、LAK（Lymphokine-activated killer cell）、巨噬细胞（macrophages）、单核细胞（monocytes）、粒细胞的培养。
TIL细胞培养明智之选
LONZA X-VIVO 15 化学成分限定无血清培养基，成分明确，不含外源生长因子，配方为TIL细胞的增殖特别优化，原产地为美国和比利时，由LONZA GMP级别的工厂生产，其X-VIVO 全系列培养基均在美国FDA备案，在国内外免疫治疗基础研究领域均具有很高的知名度，是培养TIL细胞的优先选择。
X-VIVO 15 无血清免疫细胞培养基使用方法:
Protocol #1: Medium Replacement
Approximate Time Required 2 weeks – 6 weeks
Culture Conditions:
Mammalian Cells 95% Air, 5% CO2, 35ºC-37ºC
Invertebrate Cells Air, 25ºC-27ºC
Note: Adherent cell cultures that are exposed to trypsin during the subculturing process should be converted to serum-free growth using Protocol #2.
1. Begin with cultures at maximum cell density.
2. Split cells 1:2 using serum-free medium as the diluent.
3. Incubate cells until the maximum cell density is achieved.
4. Split cells 1:5 or to 3.0x105 cells/ml for attachment independent cells or 30% confluency for attachment dependent cells using serum-free medium as the diluent.
5. Incubate cells until the maximum cell density is achieved.
6. If the cell viability is >85% at this point, and the generation time is similar to that observed with medium containing serum, the culture may be maintained on serum-free medium using a similar split schedule as originally optimized for
medium containing serum.
7. If the cells exhibit slow growth or low viability,maintain the split ratio at 1:2 or 1:5 for three successive splits. The minimum cell density should be above 3x105 cells/ml or 30% confluency during this period.
8. Gradually increase the split ratio to obtain a maximum value for the cell type being used.
Note: Some cells may require a small amount of serum for growth. If the cells have not adapted to serum-free cultivation using the above protocol, add 0.1%-0.5% serum to the culture or call Lonza Scientific Support at 1-800-521-0390
Protocol #2: Serum Dilution
Approximate Time Required 2 weeks – 6 weeks
Culture Conditions:
Mammalian Cells 95% Air, 5% CO2, 35ºC-37ºC
Invertebrate Cells Air, 25ºC-27ºC
1. Begin with cultures at maximum cell density.
2. Trypsinize adherent cultures and transfer to an appropriately sized centrifuge tube. Suspension cultures may be transferred directly to the centrifuge tube.
3. Sediment the cells using centrifugation at 350 x g for five minutes.
4. Resuspend the cells in serum-free medium containing 5% serum (V/V).
5. Adjust the cell concentration using the serumsupplemented serum-free medium to a maximum of 3x105 cells/ml for attachment independent cells or a density to achieve not less than 30% confluency.
6. Plant the cells and incubate until a maximum level of cell density is achieved.
7. Repeat steps 2-6 using a lower concentration of serum at each split. We recommend beginning at 5% serum and lowering to 2%, 1%, 0.5% and finally 0.1% prior to eliminating serum from the culture.
Note: If the culture viability drops below 80% or if the generation time increases markedly following a decrease in the serum concentration, increase the serum level to the previous value and maintain the cells for two split cycles before lowering the level of serum again. It may be necessary to institute a more gradual decline in serum concentration with these cells. Some cells may require a small amount of serum for growth. If the cells have not adapted to serum-free cultivation using the above protocol, add 0.1%-0.5% serum to the culture or call Lonza Scientific Support at 1-800-521-0390.
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——北京泽平，LONZA中国地区一级代理商