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HS-5
Tissue
骨髓、基質
Disease
正常
Synonyms
HS5

Documents

HS-5

Cell line · Product number 305451

The HS-5 cell line is a human bone marrow-derived mesenchymal stromal cell (MSC) model commonly used to study the supportive role of stromal cells in hema...

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01General information

Description

HS-5 細胞系是一種源自人類骨髓的間質基質細胞(MSC)模型,常被用於研究基質細胞在造血及癌症發展中所扮演的支持性角色。 HS-5 細胞最初是透過人類乳頭瘤病毒的 E6 和 E7 基因進行分離並永生化,其呈現成纖維細胞樣形態,並以分泌多種對造血細胞與免疫細胞相互作用至關重要的細胞因子和生長因子而聞名。 值得注意的分泌因子包括粒細胞集落刺激因子(G-CSF)、粒細胞-巨噬細胞集落刺激因子(GM-CSF)、 巨噬細胞集落刺激因子(M-CSF)、幹細胞因子(SCF)、白細胞介素-6(IL-6)、IL-8 及 IL-11 等。 這些分泌物使 HS-5 細胞能夠在體外維持造血前體細胞的生長與增殖,即使在無血清條件下亦然,這凸顯了它們在支持造血方面與原代間質幹細胞(MSCs)的功能相似性。

在腫瘤學研究中,HS-5 細胞用作研究 MSC 與癌細胞之間相互作用的模型,特別是用於研究由腫瘤微環境介導的癌細胞存活、免疫調節及藥物抗性。 研究顯示,HS-5 細胞能保護急性骨髓性白血病(AML)細胞免於因胞苷(Ara-C)和柔紅霉素(DNR)等化療藥物所誘導的凋亡,其作用機制可能涉及細胞因子訊號傳導及直接的細胞間相互作用。 HS-5 細胞的這種保護作用會增強 AML 細胞的增殖並降低化療療效,使該細胞系成為模擬骨髓微環境中藥物抗性的一項寶貴工具。此外,HS-5 細胞表達 CD73、CD90 和 CD105 等 MSC 標記物,儘管其表達水平可能與原代 MSC 有所不同。

此外,HS-5 細胞已被用於探索新型治療方法,例如測試原始石墨烯等奈米材料的細胞毒性作用。 研究顯示,HS-5 細胞暴露於石墨烯會誘發線粒體功能障礙與氧化壓力,透過增加活性氧(ROS)的產生及破壞線粒體膜電位而導致細胞凋亡,因此可作為評估奈米粒子在腫瘤微環境中與基質細胞相互作用時之毒性及治療潛力的模型。

Tissue
骨髓、基質
Disease
正常
Synonyms
  • HS5

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This human bone-marrow stromal cell line (HS-5) contains retroviral constructs encoding HPV-16 E6 and E7 oncogenes, supporting immortalization and transformation. The inserts are stably present in HS-5 cells. 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

04Identifiers

Citation
HS-5(Cytion 型號 305451)
NCBI TaxID
9606
Cellosaurus
CVCL_3720

05Expression and susceptibility

Protein expression
表達的基因:粒細胞集落刺激因子(G-CSF);粒細胞-巨噬細胞集落刺激因子(GM-CSF);巨噬細胞集落刺激因子(M-CSF); Kit 配體 (KL);巨噬細胞抑制蛋白-1α;白細胞介素-1α (IL-1α);IL-1β;IL-1RA;IL-6;IL-8;IL-11;白血病抑制因子 (LIF)
Viruses
轉化體:人類乳突病毒第16型(HPV16)。

06Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
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
Fluid renewal
每週 2 至 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

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
30年
Ethnicity
高加索人