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MC3T3-E1 Subclone 24
Tissue
骨頭

Documents

MC3T3-E1 Subclone 24

Cell line · Product number 305186

MC3T3-E1 Subclone 24 cells expressly represent a preosteoblast cell type, which plays a crucial role in bone formation. Morphologically, they exhibit a fi...

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

01General information

Description

MC3T3-E1 第 24 亞克隆細胞明確代表一種成骨前細胞類型,在骨形成過程中扮演著關鍵角色。從形態學來看,這些細胞呈現成纖維細胞樣外觀,其特徵為細長的外形及紡錘狀結構。 此特定亞克隆源自顱頂組織,該顱骨區域有助於骨形成。MC3T3-E1 亞克隆 24 細胞的一項關鍵應用在於 3D 細胞培養,研究人員可藉此在三維環境中研究這些細胞的行為與相互作用。 相較於傳統的二維細胞培養,此方法提供更貼近生理狀況的模型,有助於更深入理解骨形成過程中涉及的複雜機制。

儘管這些細胞具備眾多優勢,但必須注意其特定特性。觀察發現,當 MC3T3-E1 亞克隆 24 細胞接觸維生素 C(促進骨細胞生長的关键成分)時,其成骨細胞分化能力較差。 此外,它們無法形成礦化細胞外基質,而這是生成骨組織的關鍵步驟。MC3T3-E1 亞克隆 24 細胞的倍增時間約為 90.5 小時。

Tissue
骨頭
Applications
三維細胞培養、分化研究

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纖維細胞
Cell type
成骨細胞
Tumourigenic
是的,在免疫抑制小鼠身上

04Identifiers

Citation
MC3T3-E1 亞克隆 24(Cytion 型號 305186)
NCBI TaxID
10090
Cellosaurus
CVCL_5438

05Expression and susceptibility

Receptors expressed
副甲狀腺激素相關蛋白(PTHrP)受體
Protein expression
膠原蛋白、骨唾液蛋白(BSP)、骨鈣素(OCN)、甲狀旁腺激素(PTH)

06Handling and culture

Culture medium
Alpha MEM,含:2.0 mM 穩定谷氨醯胺,含:核苷,含:脫氧核苷,含:1.0 mM 丙酮酸鈉,含:2.2 g/L NaHCO₃,不含: 抗壞血酸(GIBCO,型號 A1049001。本公司不供應此產品;請考慮其他供應商。如需進一步協助,請告知我們。)
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。
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

Breed / subspecies
C57BL/6
Donor age
1 天