Transplantation of encapsulated cells secreting human beta-endorphin in cerebro-spinal space for cancerpain

将分泌人β-内啡肽的封装细胞移植到脑脊空间治疗癌痛

• 批准号: 07557360
• 负责人: SAITOH Youichi
• 金额: $ 0.9万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557360/
• 关键词: pain; xenograft; beta-endorphin; polymer capsule

Recently the technique of transplantation of encapsulated cells has progressed. Semipermeable membranes used for capsules allow transport of low molecular weight substances, but prevent the inward diffusion of both the humoral and cellular elements of the immune system. The proopiomelanocortin (POMC) gene transfected Neuro2A cells (mouse cell line) which secrete beta-endorphin, were enclosed in polymer capsules at a density of 5x10^6/ml, and transplanted into the spinal CSF space from the occipito-atlantal junction of male Sprague-Dawley rats. The rats with encapsulated Neuro2A showed significantly less sensitive to pain than the control animals. Morphological study revealed that the encapsulated cells survived for a month after transplantation. However, the encapsulated Neuro2A in the CSF space of Japanese monkeys showed much worse viability than that of rats. We speculated that complement or natural antibody may penetrate the capsules and attack the xenogeneic cells. Therefore, sensi … More tised sheep red blood cells were encapsulated and incubated in 10% human serum medium, and the degree of the hemolysis was measured. The result suggested that double membrane consisted of PM30 (Amicon Co.) and K6305 (Kuraray Co.) could prohibit the penetration of complement in vitro. However, Neuro2A encapsulated in double capsules showed poor viability in the CSF space of Japanese monkeys. Now its reason is studied.In our study, it has been a little difficult to control the amount of secreted opiate. However, recently, the transcriptional activation system (Tet-On system) by tetracycline has been developed, and this system may help the control of opiate secretion. The pUHD10-3-POMC and pUHD172-1neo were co-transfectred to Neuro2A,named Neuro2A-POMC.Encapsulated Neuro2A-POMC was transplanted in rat CSF space. Doxycycline 5.0,2.0,0.5 and 0 mg were injected intraperitoneally 4 times every 6 hours. The peptides secretions were well correlated with the dose of doxycycline. Tet-On system is appliable for cell therapy in CNS sytem. Less

最近，封装细胞移植技术取得了进展。用于胶囊的半透膜允许运输低分子量物质，但阻止免疫系统的体液和细胞成分向内扩散。将阿黑皮质素原（POMC）基因转染分泌β-内啡肽的Neuro2A细胞（小鼠细胞系），以5x10^6/ml的密度封装在聚合物胶囊中，并从雄性Sprague-Dawley大鼠的枕-寰椎交界处移植到脊髓CSF间隙中。与对照动物相比，包裹有 Neuro2A 的大鼠对疼痛的敏感性显着降低。形态学研究表明，封装的细胞在移植后存活了一个月。然而，封装在日本猴子脑脊液空间中的 Neuro2A 的生存能力比大鼠差得多。我们推测补体或天然抗体可能会穿透胶囊并攻击异种细胞。因此，将敏感的羊红细胞封装在10%人血清培养基中培养，并测量溶血程度。结果表明，PM30（Amicon Co.）和K6305（Kuraray Co.）组成的双层膜可以在体外抑制补体的渗透。然而，包裹在双胶囊中的 Neuro2A 在日本猴的脑脊液空间中表现出较差的活力。现在对其原因进行了研究。在我们的研究中，控制阿片类物质的分泌量有点困难。然而，最近，四环素转录激活系统（Tet-On系统）已被开发出来，该系统可能有助于控制阿片类药物的分泌。将pUHD10-3-POMC和pUHD172-1neo共转染到Neuro2A中，命名为Neuro2A-POMC。将封装的Neuro2A-POMC移植到大鼠脑脊液空间中。每6小时腹腔注射强力霉素5.0、2.0、0.5和0mg，共4次。肽的分泌与多西环素的剂量密切相关。 Tet-On系统适用于中枢神经系统细胞治疗。较少的

项目成果

Y.Saitoh et al: "Analgesia induced by trasplantation of encapsulated tumor cells secreting beta-endorphin" J Neurosurg. 82. 630-634 (1995)

Y.Saitoh 等人：“分泌 β-内啡肽的封装肿瘤细胞移植诱导的镇痛”J Neurosurg。

S.Hiraga et al: "Transformation of type 1 astrocytes with N-Ethyl-N-Nitrosourea : Establishment of an in vitro system and the role of the p53 gene" Glia. 13. 51-63 (1995)

S.Hiraga 等人：“用 N-乙基-N-亚硝基脲转化 1 型星形胶质细胞：体外系统的建立和 p53 基因的作用”神经胶质细胞。

H.Hayashi et al: "Long survival of xenografted bioartificial pancreas with a mesh-reinforced polyvinyl alcohol hydrogel bag employing a B-cell line (MIN6)" Transplantation Proceed. 28. 1428-1429 (1996)

H.Hayashi 等人：“采用 B 细胞系 (MIN6) 的网状强化聚乙烯醇水凝胶袋异种移植生物人工胰腺的长期存活”进行移植。

A C Gore,Y Saitoh et al.: "Effects of adrenal medulla transplantation into the third ventricle on the onset of puberty in female rhesus monkeys." Experimental Neurology. 140. 172-183 (1996)

A C Gore、Y Saitoh 等人：“肾上腺髓质移植到第三脑室对雌性恒河猴青春期开始的影响”。

Youichi Saitoh et al: "Analgesia induced by transplantation of encapsulated tumor cells secreting β-endorphin" Analgesic Digest. 2. 18-21 (1995)

Youichi Saitoh 等：“分泌β-内啡肽的封装肿瘤细胞移植诱导的镇痛”镇痛文摘。