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WiDr
Tissue
结肠
Disease
腺癌
Synonyms
WiDR, WIDR, WiDr/S, WiDr-TC, WiDrTC, LED-WiDr, Led-WiDr

Documents

WiDr

Cell line · Product number 300377

According to Chen TR, 1987, the WiDr cell line is a derivative of the HT-29 cell line. The cells are negative for Colon Antigen 3 expression, but positive fo...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
1

01General information

Description
根据 Chen TR，1987 年，WiDr 细胞系是 HT-29 细胞系的衍生物。这种细胞的结肠抗原 3 表达阴性，但通过免疫过氧化物酶染色，角蛋白表达阳性。细胞表达 p53 抗原（产生的 p53 在 273 位发生 G -> A 突变，导致 Arg -> His）。WiDr 细胞的生长受到肿瘤坏死因子 alpha 的抑制（TNF 抑制剂二氢叶酸还原酶对 WiDr 细胞有很强的细胞毒性）。
Tissue
结肠
Disease
腺癌
Synonyms
  • WiDR
  • WIDR
  • WiDr/S
  • WiDr-TC
  • WiDrTC
  • LED-WiDr
  • Led-WiDr

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是，在裸鼠身上

04Identifiers

Citation
WiDr（Cytion 目录号 300377）
NCBI TaxID
9606
Cellosaurus
CVCL_2760

05Expression and susceptibility

Receptors expressed
表皮生长因子（EGF）
Protein expression
癌胚抗原阳性
Antigen expression
HLA A24、A32、B15、B18
Isoenzymes
PGM1，1-2，PGM3，1-2，G6PD，B，ES-D，1，PEP-D，1，6PGD，A
Products
癌胚抗原 (CEA) 118 纳克/106 个细胞/10 天、结肠特异性抗原 (CSAp)、转化生长因子 beta、角蛋白

06Handling and culture

Culture medium
DMEM:Ham's F12（1:1），w: 3.1 g/L 葡萄糖，w: 2.5 mM L-谷氨酰胺，w: 15 mM HEPES，w: 0.5 mM 丙酮酸钠，w: 1.2 g/L NaHCO3（Cytion 文章编号 820400a）。
Incubation atmosphere

37°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
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 1 至 2 次
Post-thaw recovery
解冻后，将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中，使细胞从冷冻过程中恢复并至少贴壁24小时。
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

Donor age
44 年