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MH7A
Tissue
滑膜
Disease
滑膜成纖維細胞(經 SV40 永生化;模擬類風濕性關節炎滑膜細胞)
Metastatic site
不適用(非致瘤性滑膜成纖維細胞模型)

Documents

MH7A

Cell line · Product number 305036

MH7A is a human synovial fibroblast cell line immortalized by stable SV40 large T antigen expression, established from synovial tissue of a female Japanes...

US$800.00excl. tax and shipping
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  • CoA
1

01General information

Description

MH7A 是一種透過穩定表達 SV40 大 T 抗原而永生化的人類滑膜成纖維細胞系,源自一名日本女性患者的滑膜組織。 MH7A 以貼壁培養方式生長,呈現成纖維細胞樣形態,並保留類風濕性關節炎滑膜成纖維細胞(RASFs)的關鍵功能特徵,包括對促炎性細胞因子(IL-1β、 TNF-α)的反應性、基質金屬蛋白酶(MMPs)的表達,以及侵襲軟骨基質的能力。它是研究滑膜細胞生物學及類風濕性關節炎病理機制最廣泛使用的體外模型之一。

MH7A 適用於類風濕性關節炎研究、滑膜細胞活化、MMP 與 TIMP 調控、細胞因子訊號傳導(NF-κB、JAK-STAT、MAPK)、抗風濕藥物評估(DMARDs、生物製劑、JAK 抑制劑),以及關節破壞的共培養模型。 該細胞系提供了一種一致且可重複的類風濕性關節炎滑膜細胞模型,可避免原代患者來源類風濕性關節炎滑膜成纖維細胞(RASFs)所存在的變異性及供應有限的問題。

Tissue
滑膜
Disease
滑膜成纖維細胞(經 SV40 永生化;模擬類風濕性關節炎滑膜細胞)
Metastatic site
不適用(非致瘤性滑膜成纖維細胞模型)
Applications
類風濕性關節炎研究;滑膜細胞活化;MMP/TIMP 調控;NF-κB/JAK-STAT/MAPK 訊號傳導;抗風濕藥物評估;關節破壞共培養模型;細胞因子訊號傳導

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: MH7A was immortalized by stable integration of SV40 large T antigen. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
成纖維細胞樣
Cell type
成纖維細胞樣滑膜細胞
Doubling time
約 24 至 36 小時

04Identifiers

Citation
MH7A(Cytion 型號 305036)
NCBI TaxID
9606
Cellosaurus
CVCL_0427

05Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
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,5 分鐘,37°C
Subculturing
移除培養基,用不含鈣和鎂的PBS清洗,加入0.25%胰蛋白酶浸沒細胞,於37°C孵育3–5分鐘,重新懸浮於培養基中,以300×g離心3分鐘,棄去上清液,重新接種。
Split ratio
1:8 的比例
Seeding density
1 至 3 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 2 至 5 × 10⁴ 個細胞/cm² 的密度接種至培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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.

06Quality control and cryopreservation

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。

07Donor

Donor age
年齡不詳
Ethnicity
日文