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T84
Tissue
冒號
Disease
癌
Metastatic site
肺
Synonyms
T-84, T 84

Documents

T84

Cell line · Product number 300354

This line exhibits tight junctions, and desmosomes between adjacent cells. The cells should be maintained at high density (at least 1/4 confluency).

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

01General information

Description
此細胞系呈現緊密連接,且相鄰細胞之間存在橋粒。應將細胞維持在高密度狀態(至少達到 1/4 融合度)。
Tissue
冒號
Disease
癌
Metastatic site
肺
Applications
大腸直腸癌研究;腸道上皮生物學;緊密連接與屏障功能研究;結腸運輸生理學;囊性纖維化跨膜傳導調節蛋白(CFTR)研究;藥物吸收與代謝;異種移植模型
Synonyms
  • T-84
  • T 84

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
GMO status
Non-GMO
GMO status details
No genetic modification; wildtype colon carcinoma cell line (KRAS G13D heterozygous mutation is an endogenous somatic change, not a genetic engineering modification)
Biosafety level
1

03Characteristics

Morphology
上皮樣
Cell type
上皮細胞
Tumourigenic
是的,在裸鼠身上
Karyotype
幹系模式染色體數為 56,出現率為 28%,多倍體比例為 12.4%。 在所檢視的大多數中期細胞中,有十八個標記是共通的。正常X染色體及第13號染色體均缺失,第2、4及22號染色體為單拷貝,而第12號染色體則為4拷貝。透過Q帶觀察未檢測到Y染色體。近50%的細胞出現雙極性分裂(DM)。
Doubling time
約 48 至 72 小時

04Identifiers

Citation
T84(Cytion 型號 300354)
NCBI TaxID
9606
Cellosaurus
CVCL_0555

05Expression and susceptibility

Receptors expressed
肽類激素、神經傳導物質
Antigen expression
角蛋白 +(免疫過氧化物酶染色)
Isoenzymes
G6PD、B、PGM1、1、PGM3、1、ES-D、1、Me-2、1-2、AK-1、1、GLO-1、1-2
Mutational profile
T84 細胞攜帶第 13 個密碼子處的 Kras 雜合突變:GGC(野生型甘胺酸)>GAC(天冬氨酸)
Products
癌胚抗原(CEA),每 10 exp6 個細胞每 10 天 600 ng/ml,角蛋白

06Handling and culture

Culture medium
Ham's F12,含:1.0 mM 穩定型谷氨醯胺、1.0 mM 丙酮酸鈉、1.1 g/L NaHCO₃(Cytion 產品編號 820600a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% 的胎牛血清(FBS)
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
Split ratio
1 至 3
Seeding density
1 至 2 × 10⁴ 個細胞/cm²(需維持至少 1/4 的匯合度,以保留緊密連接的表型)
Fluid renewal
每週 2 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並靜置至少 24–48 小時以利細胞貼壁。應維持高密度細胞狀態(≥25% 貼壁率),以確保緊密連接的形成。
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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

08Donor

Donor age
72年
Ethnicity
未指定族裔