Kang Seok-Gu, Jeun Sin Soo, Lim Jung Yeon, Yoo Do Sung, Huh Pil Woo, Cho Kyung Souk, Kim Dal Soo, Shin Hyung-Jin, Kim Jong Hyun, Kim Moon Chan, Kang Joon Ki
Department of Neurosurgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Republic of Korea.
Childs Nerv Syst. 2005 Jul;21(7):528-38. doi: 10.1007/s00381-005-1216-3. Epub 2005 Jun 3.
Marrow stromal cells (MSCs) have been shown to have the capacity of orthodox and unorthodox plasticity. In this study, the authors tried to access in vitro cytotoxicity of MSCs from rat and also to differentiate MSCs into immune effector cell.
Rat MSCs (rMSCs) were isolated by standard methodology and were activated by interleukin-2 (IL-2), interleukin-15 (IL-15), granulocyte macrophage colony stimulating factor, and combinations, which were effector cells. Cytotoxicity of rMSCs and activated rMSCs against the target cells (9L rat glioma cell line) was estimated using visual survival cell assay. Phenotypes of these various activated cells were determined using flow cytometry. The secreted protein from effector cells was estimated by enzyme-linked immunosorbent assay. The expression of immune response-related genes in activated cells was measured.
There was a significant cytotoxicity of rMSCs activated with various cytokine combinations. After various cytokine activations of rMSCs, the population of immune effector cells (CD8, CD161a) and immune reaction-related proteins (IL-4, gamma-INF) might increase. Apoptosis may be one of the lysis mechanisms of target cells by activated rMSCs. The contributing genes could be gamma-INF, FasL, and perforin.
This study suggests that rMSC may be used as adoptive transfer therapy in patients suffering from malignant brain tumor, but we have to investigate orthotopic animal study for the proper translation.
骨髓基质细胞（MSCs）已被证明具有常规和非常规可塑性。在本研究中，作者试图评估大鼠MSCs的体外细胞毒性，并将MSCs分化为免疫效应细胞。
采用标准方法分离大鼠MSCs（rMSCs），并用白细胞介素-2（IL-2）、白细胞介素-15（IL-15）、粒细胞巨噬细胞集落刺激因子及其组合进行激活，这些均为效应细胞。使用视觉存活细胞测定法评估rMSCs和活化rMSCs对靶细胞（9L大鼠胶质瘤细胞系）的细胞毒性。使用流式细胞术确定这些不同活化细胞的表型。通过酶联免疫吸附测定法评估效应细胞分泌的蛋白质。测量活化细胞中免疫反应相关基因的表达。
用各种细胞因子组合激活的rMSCs具有显著的细胞毒性。rMSCs经各种细胞因子激活后，免疫效应细胞（CD8、CD161a）群体和免疫反应相关蛋白（IL-4、γ-干扰素）可能增加。凋亡可能是活化rMSCs裂解靶细胞的机制之一。相关基因可能是γ-干扰素、FasL和穿孔素。
本研究表明，rMSC可用于恶性脑肿瘤患者的过继性细胞转移治疗，但我们必须进行原位动物研究以实现恰当转化。
