Li Qiu-bai, Tang Xiao-qiong, Zhao Zhi-gang, Wang Hong-xiang, You Yong, Chen Zhi-chao, Zou Ping
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Zhonghua Yi Xue Za Zhi. 2007 Mar 27;87(12):812-5.
To study the protective role of amifostine (WR-2721) in the mesenchymal stem cells (MSC) derived from non-Hodgkin lymphoma (NHL) bone marrow treated with etoposide (VP-16).
MSC were obtained from non-Hodgkin lymphoma bone marrow, cultured in expanded medium, and then divided into 4 groups: Group A, without treatment by either WR-2721 or VP-16 and used as control group, Group B, treated with WR-2721, Group C, treated with WR-2721 + VP-16, and Group D, treated with VP-16 alone. Inverted microscopy was used to observe the growth of the MSC. Flow cytometry was used to observe the apoptosis and immunophenotypes of the MSC. Mononuclear growth and apoptosis of the MSC Mononuclear bone marrow cells were obtained from 5 healthy volunteers and added into the MSC of the 4 groups, 4 weeks later methylcellulose progenitor assay and then inverted microscopy were used to measure the numbers of colony so as to detect the hematopoiesis. MSC of different groups were put into osteocyte-inducing and adipocyte-inducing media respectively, and 2 weeks later Von Kossa staining and oil-red O staining were used to identify the differentiation into bone and fat.
The NHL derived MSC of the 4 groups all showed a typical fibroblast-like morphology and were all positive in CD29, CD44, and CD105, while negative in CD11b, CD31, CD34, CD45, and HLA-DR. The apoptotic rate of Group D was 31.2% +/- 4.3%, significantly higher than those of the other 3 groups (all P < 0.05), and the apoptotic rate of Group C was significantly higher than those of Groups A and B (both P < 0.05), however, still significantly lower than that of Group D (P < 0.05). The ability to support hematopoiesis of Group B was not significantly different from that of Group A, and Groups C and D showed a lower ability to support hematopoiesis in comparison with Groups A and B, However, the ability of Group C was still significantly higher than that of Group D (P < 0.05). Under suitable conditions, all the MSC differentiated into osteocytes or adipocytes.
Amifostine alone has no effects on proliferation, apoptosis, immunophenotype or ability of hematopoiesis support of the MSC in vitro; however, it can protect NHL patient-derived MSC from injury by VP-16 in vitro.
研究氨磷汀（WR-2721）对依托泊苷（VP-16）处理的非霍奇金淋巴瘤（NHL）骨髓间充质干细胞（MSC）的保护作用。
从非霍奇金淋巴瘤骨髓中获取MSC，在扩增培养基中培养，然后分为4组：A组，未用WR-2721或VP-16处理，作为对照组；B组，用WR-2721处理；C组，用WR-2721 + VP-16处理；D组，仅用VP-16处理。采用倒置显微镜观察MSC的生长情况。采用流式细胞术观察MSC的凋亡及免疫表型。MSC单核细胞的生长与凋亡 从5名健康志愿者获取单核骨髓细胞并加入4组MSC中，4周后进行甲基纤维素祖细胞测定，然后用倒置显微镜测量集落数量以检测造血功能。将不同组的MSC分别置于成骨细胞诱导培养基和脂肪细胞诱导培养基中，2周后采用冯库萨染色和油红O染色鉴定其向骨和脂肪的分化情况。
4组NHL来源的MSC均呈现典型的成纤维细胞样形态，CD29、CD44和CD105均呈阳性，而CD11b、CD31、CD34、CD45和HLA-DR均呈阴性。D组的凋亡率为31.2%±4.3%，显著高于其他3组（均P < 0.05），C组的凋亡率显著高于A组和B组（均P < 0.05），但仍显著低于D组（P < 0.05）。B组支持造血的能力与A组无显著差异，C组和D组支持造血的能力低于A组和B组，然而，C组的能力仍显著高于D组（P < 0.05）。在合适条件下，所有MSC均分化为成骨细胞或脂肪细胞。
单独使用氨磷汀对体外培养的MSC的增殖、凋亡、免疫表型或支持造血的能力无影响；然而，它可在体外保护NHL患者来源 的MSC免受VP-16的损伤。
