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MDS-L
Tissue
骨髓
Disease
骨髓增生异常综合征
Synonyms
MDSL

Documents

MDS-L

Cell line · Product number 305826

MDS-L is a human myelodysplastic syndrome (MDS)-derived cell line originally established from the MDS92 cell line, which itself was derived from the bone ...

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01General information

Description

MDS-L系人骨髓增生异常综合征(MDS)衍生的细胞系,最初由MDS92细胞系建立,而MDS92细胞系本身源自一名具有del(5q)染色体异常的MDS患者骨髓。 MDS92细胞系包含处于不同分化阶段的异质性髓系细胞混合体,而MDS-L则代表一种具有更统一特征的原始细胞亚系,其特征为未成熟的髓系祖细胞。 MDS-L在体外增殖仍依赖白细胞介素-3(IL-3),其细胞因子敏感性与原代MDS前体细胞一致。该细胞系携带多重遗传改变,包括TP53纯合突变及NRAS和CEBPA基因获得性突变。这些改变共同反映了高危MDS典型的克隆演变与白血病转化潜能。

MDS-L细胞系已被广泛用作研究MDS发病机制、分化阻滞及治疗耐药性分子机制的模型。通过该细胞系的重要发现之一是:经逆转录病毒转导强制表达粒细胞集落刺激因子受体(G-CSFR)后,在G-CSF刺激下可实现粒细胞分化。 该现象通过形态学改变、CD11b表达增强及硝基蓝四唑(NBT)还原活性提升得到证实——这些均是粒细胞终末成熟的标志。这些结果揭示了MDS-L在恢复相应信号组分后具有内在分化能力,为针对MDS分化缺陷的潜在基因治疗方案提供了新思路。

除遗传学与功能研究外,MDS-L细胞系在阐明组蛋白修饰对疾病进展的作用方面发挥了关键作用。 值得注意的是,在MDS-L中发现了通常与小儿胶质瘤相关、但在血液恶性肿瘤中罕见的组蛋白H3-K27M突变,该突变被证实可抑制EZH2介导的组蛋白甲基化。这种表观遗传改变导致H3-K27甲基化水平普遍降低,并与p16等抑癌基因表达改变相关。 通过差异化IL-3培养条件获得的含/不含该突变的MDS-L亚系,进一步揭示了MDS内部的表观遗传异质性及其对IL-3依赖性生长和治疗反应的影响。这些独特特性使MDS-L成为研究MDS分子演变、治疗靶点及其向急性髓系白血病转化的强大体外和体内模型。

Tissue
骨髓
Disease
骨髓增生异常综合征
Synonyms
  • MDSL

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
1

03Identifiers

Citation
MDS-L(赛迪昂产品编号 305826)
NCBI TaxID
9606
Cellosaurus
CVCL_A8QV

04Expression and susceptibility

Mutational profile
突变:CEBPA,简单突变,p.Gln311Ter (c.931C>T),杂合子,H3C3,简单突变,p.Lys28Met (c.83A>T),杂合子,NRAS,简单突变, p.Gly12Ala (c.35G>C),杂合子,TP53,单点突变,c.672+1G>A,纯合子,注=剪接供体突变

05Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
向培养基中添加10%胎牛血清和20 ng/ml人重组IL-3。
Dissociation reagent
无
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

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。

07Donor

Donor age
52 年
Ethnicity
日语