Isolation and identification of cancer stem cells from human osteosarcom by serum-free three-dimensional culture combined with anticancer drugs
Isolation and identification of cancer stem cells from human osteosarcom by serum-free three-dimensional culture combined with anticancer drugs
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无血清三维培养联合抗癌药物分离鉴定人骨肉瘤干细胞
DOI：
10.1007/s11596-010-0114-4
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发表时间：
2010-02
期刊：
影响因子：
--
通讯作者：
Xiao, Jun
中科院分区：
文献类型：
--
作者：
Xiong, Wei;Zhou, Song;Niu, Pengyan;Fang, Zhong;Li, Feng;Chen, Wenjian;Xiao, Jun
采用无血清三维培养结合抗癌药物的方法从人骨肉瘤中分离并鉴定肿瘤干细胞。将人骨肉瘤原代细胞用胰蛋白酶消化，制成单细胞悬液，均匀混合于1.2%海藻酸钠凝胶中。用无血清DMEM/F12培养基培养单细胞藻酸盐凝胶。向培养基中加入依匹林（0.8 μg/mL）以富集CSC。常规培养7 ~ 10 d后，大部分藻酸盐凝胶中的细胞被表射杀死。但很少有细胞存活，并形成了一些单细胞克隆球。对Oct 3/4和Nanog进行免疫荧光染色，以发现具有自我更新和多潜能分化特性的细胞。将克隆球中的细胞移植到BALB/c小鼠体内，检测其体内致瘤性。结果显示，在单细胞克隆球中发现了部分Oct 3/4（TRITC）和Nanog（TRITC）阳性细胞，且阳性细胞主要集中在球的核心。球形细胞在小鼠体内可形成骨肉瘤。结论：单细胞克隆球培养的细胞具有表达部分干细胞基因（Oct 3/4和Nanog）、抗肿瘤药物和体内致瘤性的特性。综上所述，认为骨肉瘤无血清三维培养结合抗癌药物获得的细胞为肿瘤干细胞。
The cancer stem cells (CSCs) from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified. The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension, and mixed homogeneously into 1.2% alginate gel. Single-cell alginate gel was cultured with serum-free DMEM/F12 medium. Epirubicin (0.8 μg/mL) was added to the medium to enrich CSCs. After cultured conventionally for 7 to 10 days, most of cells suspended in alginate gel were killed by epirubicin. But few cells survived and some single-cell cloning spheres formed. Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation. Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicityin vivo. The results showed that some cells positive for Oct3/4 (TRITC) and Nanog (TRITC) were found in single-cell cloning spheres, and most of positive cells were concentrated in the core of sphere. Cells from spheres could form osteosarcoma in the body of mice. It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes (Oct3/4 and Nanog), resisting anti-cancer drugs, and tumorigenicityin vivo. To sum up, it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.
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影响因子：
3.8
作者：
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通讯作者：
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作者：
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作者：
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通讯作者：
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