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SVI
Tissue
肾脏

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SVI

Cell line · Product number 400495

The SVI cell line has been cloned from the outgrowth of glomeruli which were isolated from H-2kb-tsA58 transgenic mice. The mice carry a temperature-sensitiv...

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1

01General information

Description
SVI 细胞系是从从 H-2kb-tsA58 转基因小鼠体内分离出来的肾小球外生组织中克隆出来的。这种小鼠携带对温度敏感的 SV40 大 T 抗原变体，受 IFN 诱导的 H-2kb 启动子控制。细胞在 33 摄氏度下增殖，在 37 摄氏度下分化。目前，该细胞已成功培养了 40 多次，未发现表型变化。在形态和几种标记物的表达方面，SVI 与 E11 非常相似。例如，podocin 和 WT1 的表达量低于 E11。分化：开始分化过程，将非融合烧瓶放入 38 摄氏度/5% CO2 的培养箱中至少培养 14 天，以完成分化。无需添加干扰素-γ (INF-γ)。
Tissue
肾脏

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine podocyte cell line (SVI) contains a conditionally active SV40 Large T-Antigen transgene as part of the ImmortoMouse model, supporting temperature-sensitive immortalization. The construct is stably present in podocyte-derived cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Cell type
荚膜细胞

04Identifiers

Citation
SVI（Cytion 目录编号 400495）
NCBI TaxID
10090
Cellosaurus
CVCL_5943

05Expression and susceptibility

Protein expression
WT1、Lmx1b、肾素、NEPHI、FAT、P-cadherin、CD2AP、ZO-I、podocalyxin、podoplanin、synpo、podocin、TRPC6 和 GAPDH。

06Handling and culture

Culture medium
RPMI 1640，w：2.0 mM 稳定谷氨酰胺，w：2.0 g/L NaHCO3（Cytion 文章编号 820700a）。
Incubation atmosphere

33°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基，用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS，T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞，T25 烧瓶用 1-2 毫升，T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟，使其分离。孵育后，用 10 毫升培养基轻轻混合细胞使其重悬，然后用 300xg 离心 3 分钟。弃去上清液，用新鲜培养基重悬细胞，并将其转移到已装有新鲜培养基的新烧瓶中。
Seeding density
接种T75细胞培养瓶，细胞密度为1×10⁴个细胞/cm²（约60,000个细胞/ml，单个T75瓶含12ml培养基），进行增殖培养。将细胞置于33℃/5% CO₂环境中培养，直至细胞瓶达到约75%融合度。
Fluid renewal
每周 3 次
Thawing and culturing

  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染，每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基（包括 FBS）+10% DMSO 或 CM-1（Cytion 商品目录编号 800100）作为冷冻保存培养基，以获得足够的解冻后存活率，CM-1（Cytion 商品目录编号 800100）含有优化的渗透保护剂和代谢稳定剂，可提高恢复能力并减少冷冻引起的应激。
Storage conditions

如需长期保存，可将样品瓶放入气相液氮中，温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运，并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后，应立即检查容器，并立即将小瓶转移到适当的储藏室。

08Donor

Breed / subspecies
(CBA/Ca x C57BL/10)Tg(H2KbtsA58) Immort
Donor age
成人