中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (19): 3024-3029.doi: 10.12307/2022.381

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓间充质干细胞条件培养液对多发性骨髓瘤细胞增殖的影响

黄  美1，王飞清2，梁惠玲2，杨  姁2，赵嘉宁2，王  琨2，刘燕青2，周  媛2，王季石1，3，李艳菊1，3， 刘  洋2，4    

  1. 1贵州医科大学，贵州省贵阳市   550004；2贵州中医药大学第一附属医院，贵州省贵阳市   550001；3贵州医科大学附属医院血液科，贵州省贵阳市   550004；4贵州医科大学细胞工程生物医药技术国家地方联合工程实验室，贵州省贵阳市   550004
  • 收稿日期:2021-06-01 修回日期:2021-07-24 接受日期:2021-09-01 出版日期:2022-07-08 发布日期:2021-12-28
  • 通讯作者: 李艳菊，博士，副主任医师，贵州医科大学附属医院血液科，贵州省贵阳市 550004 刘洋，博士，副教授，贵州医科大学细胞工程生物医药技术国家地方联合工程实验室，贵州省贵阳市 550004
  • 作者简介:黄美，女，1995年生，贵州省贵阳市人，汉族，贵州医科大学在读硕士，主要从事干细胞组织工程研究。
  • 基金资助:
    国家自然科学基金(31660326)，项目负责人：刘洋；中国博士后科学基金(2018M640938)，项目负责人：刘洋；中国博士后科学基金(43XB3794XB)，项目负责人：李艳菊；贵州省科学技术厅项目(黔科合基础[2016]1019)，项目负责人：刘洋；贵州省科学技术厅项目(黔科合LH字[2016]7505)，项目负责人：刘燕青；贵州省科学技术厅项目(黔科合LH字[2017]7140号)，项目负责人：王飞清；贵州省教育厅拔尖人才项目(黔教合 KY[2016]074)，项目负责人：刘洋；贵州省中医药管理局项目(QZYY-2018-062)，项目负责人：周媛；贵中医科院内[2019]45号，项目负责人：王琨

Effect of bone marrow mesenchymal stem cell conditioned medium on the proliferation of multiple myeloma cells

Huang Mei1, Wang Feiqing2, Liang Huiling2, Yang Xu2, Zhao Jianing2, Wang Kun2, Liu Yanqing2, Zhou Yuan2, Wang Jishi1, 3, Li Yanju1, 3, Liu Yang2, 4   

  1. 1Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou Province, China; 3Department of Hematology, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 4National & Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Received:2021-06-01 Revised:2021-07-24 Accepted:2021-09-01 Online:2022-07-08 Published:2021-12-28
  • Contact: Li Yanju, MD, Associate chief physician, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Department of Hematology, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China Liu Yang, MD, Associate professor, First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550001, Guizhou Province, China; National & Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • About author:Huang Mei, Master candidate, Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 31660326 (to LY); China Postdoctoral Science Foundation, No. 2018M640938 (to LY), No. 43XB3794XB (to LYJ); the grant from the Science and Technology Department of Guizhou Province, No. [2016]1019 (to LY), No. LH[2016]7505 (to LYQ), No. LH[2017]7140 (to WFQ); the Top Talent Program of Guizhou Provincial Department of Education, No. KY[2016]074 (to LY); the Project Foundation of Guizhou Administration of Traditional Chinese Medicine, No. QZYY-2018-062 (to ZY); the grant from the Guizhou University of Traditional Chinese Medicine, No. [2019]45 (to WK)

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摘要：

文题释义：
骨髓间充质干细胞条件培养液：骨髓间充质干细胞贴壁生长过程中会分泌各种生长因子、细胞因子等多种活性物质于培养液中，收集骨髓间充质干细胞上清培养基，经过滤和超浓缩处理后，得到含有该间充质干细胞分泌物的培养液，称之为骨髓间充质干细胞条件培养液。
人骨髓间充质干细胞条件培养液成分和作用：骨髓间充质干细胞的旁分泌作用包括趋化性能、免疫调节功能、细胞生长支持功能、抗细胞凋亡及血管生成功能等，是通过其分泌的胰岛素样生长因子1、血管内皮生长因子、转化生长因子β1、肝细胞生长因子等细胞因子实现的。

背景：近年来，骨髓微环境与多发性骨髓瘤细胞生长、存活及耐药性的关系成为科研人员研究热点，骨髓间充质干细胞条件培养液干预多发性骨髓瘤细胞株的实验鲜有报道。
目的：探讨骨髓间充质干细胞条件培养液对多发性骨髓瘤细胞增殖的影响。
方法：经Ficoll密度梯度离心法及贴壁纯化健康供者骨髓来源间充质干细胞，收集第3-5代骨髓间充质干细胞的上清培养基，超滤离心管制备浓缩骨髓间充质干细胞条件培养液，制备含10%人骨髓间充质干细胞条件培养液的1640完全培养基，诱导多发性骨髓瘤RPMI8226和U266细胞株培养48 h，MTT法、EdU荧光染色检测细胞增殖能力；流式细胞仪和PI荧光染色检测细胞凋亡情况；Real-time PCR和Western blot检测细胞中BCL-2、BTK mRNA、蛋白表达水平。
结果与结论：骨髓间充质干细胞条件培养液能促进多发性骨髓瘤细胞株的增殖(P < 0.05)，抑制多发性骨髓瘤细胞的凋亡(P < 0.05)，与BCL-2、BTK的mRNA和蛋白高表达密切相关(P < 0.05)。结果表明，骨髓间充质干细胞条件培养液可促进多发性骨髓瘤细胞增殖。

https://orcid.org/0000-0001-6037-9727 (刘洋) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 骨髓间充质干细胞, 多发性骨髓瘤, 条件培养液, 增殖, 凋亡

Abstract: BACKGROUND: In recent years, relationship between the bone marrow microenvironment and the growth, survival and drug resistance of multiple myeloma cells has become a research hotspot for researchers. Few studies have evaluated the effects of bone marrow mesenchymal stem cells conditioned culture medium on multiple myeloma cell lines.  
OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cell conditioned medium on the proliferation of multiple myeloma cell lines.
METHODS:  Ficoll density gradient centrifugation and adherent method were used to purify bone marrow mesenchymal stem cells from healthy donors. The supernatants of 3-5 generations of bone marrow mesenchymal stem cells were collected in the culture medium. Ultrafiltration centrifuge tube was used to prepare bone marrow mesenchymal stem cell conditioned medium and to prepare 1640 complete medium containing 10% human bone marrow mesenchymal stem cell conditioned medium so as to induce the culture of multiple myeloma cell lines RPMI8226 and U266 for 48 hours. Cell proliferation was detected by MTT assay and EdU fluorescence staining. Cell apoptosis was detected by flow cytometry and PI fluorescence staining. Real-time PCR and western-blot assay were used to detect mRNA and protein levels of multiple myeloma cell lines BCL-2 and BTK.  
RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cell conditioned medium could promote the proliferation of multiple myeloma cell lines (P < 0.05), reduce the apoptosis rate of cancer cells (P < 0.05) and was associated with high mRNA and protein expression of BCL-2 and BTK (P < 0.05). The results showed that the bone marrow mesenchymal stem cell conditioned medium promoted the proliferation multiple myeloma cell lines.

Key words: ">bone marrow mesenchymal stem cells, multiple myeloma, conditioned medium, proliferation, apoptosis

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