急性髓性白血病细胞体外培养及其生物学特性的研究

Biological characteristics of acute myeloid leukemia after culture in vitro

目的:研究急性白血病细胞体外培养的方法及其在体外培养过程中生物学特性的变化。方法:分别收集10例急性髓性白血病细胞,用含细胞因子无血清液体培养基进行培养2～4周,并对培养前后细胞进行细胞计数,流式细胞仪检测CD33、CD13、CD34及CD14表面抗原鉴定细胞分化,半固体培养法对培养前后的白血病细胞进行集落形成能力检测。结果:含细胞因子无血清液体培养基能有效支持AML细胞进行体外短期培养及增殖,14d时细胞得到明显增殖,达(25.1±11.7)倍,优于培养前,P<0.05;培养28d时细胞数较前明显减少,P<0.05。集落形成单位培养14d后与培养前比较差异无统计学意义,P>0.05;但培养28d后显著低于培养前,P<0.05。在体外悬浮培养14d时,CD33、CD13、CD34及CD14的表达率与培养前差异无统计学意义(P>0.05),但至28d时,CD33、CD13及CD14的表达率高于培养前,而CD34表达率比培养前降低,P<0.05。结论:含细胞因子无血清液体培养基能有效支持急性髓性白血病细胞短期体外培养并维持其生物学特性。

OBJECTIVE： To establish a system for the culture of acute myeloid leukemia （AML） and study the acute myeloid leukemia cells biology after culture in vitro. METHODS.. Ten cases of acute myeloid leukemia cells were collected and cultured in a serumfree culture system supplemented with cytokines for 2 to 4 weeks. CD33, CD13, CD34 and CD14 positive ceils were monitored by FACS. The colony forming unit （CFU） was assessed by a semisolid culture assay. RESULTS： The culture system used in this study supported the short-term culture and proliferation of acute myeloid leukaemia cells. The maximum expansion fold of the total cells was achieved at day 14, which was 25.1±11.7 （P〈0.05）. The total cell number decreased significantly after 28 days culture （P〈0.05）. CFU number maintained for 14 days after culture, but after 28 days, the culture CFU number decreased significantly （P〈 0.05）. The expressions of CD33, CD13, CD34 and CD14 also maintained for 14 days after culture, but the expressions of CD33, CD13 and CD14 increased and CD34 reduced as compared with day 0 （P〈0. 05） after 28 days＇ culture. CONCLUSIONS： The serum free system supplemented with cytokines can effectively support the culture and expansion of AML cells for short-term culture, meanwhile, the biological characteristics of the leukemia cells can be maintained.