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Celltherapy for pain with the transplantation of encapsulated xenogeneic cells secreting opiate

通过移植分泌阿片剂的封装异种细胞来治疗疼痛的细胞疗法

基本信息

批准号：
06807112
负责人：
SAITOH Youichi
金额：
$ 1.28万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06807112/
关键词：
pain xenograft beta-endorphin polymer capsule

项目摘要

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 the cellular elements of the immune system. The proopiomelanocortin gene transfected Neuro2A which secrete beta-endorphin, were enclosed in polymer capsules at a density of 5*10^6/ml, and transplanted into the spinal CSF space from the occipto-atlantal junction of male Sprague-Dawley rats. Three analgesiometric tests-the tail pinch test, the hot plate test, and electrical stimulation test-showed that the rats with encapsulated Neuro2A were significantly less sensitive to pain after transplantation than control animals. Morphological study revealed that the encapsulated cells survived for a month after transplantation into the CSF space. 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, encapsulated sensitised sheep red blood cells were incubated in 10% human serum medium. To assess the degree of the hemolysis, absorbance at the wavelength (541nm) of medium was measured. The result suggested that double membrane consisted of PM30 (Amicon Co. ; MW cut off of 30kD) and K6305 (Kuraray Co. ; MW cut off of 13kD) could prohibit the penetration of complement. Next we will try the transplantation of double membrane capsules containing Neuro2A in the CSF space of Japanese monkey.In our cell therapy, it has been a little difficult to control the amount of secreted opiate. However, recently, the transcriptional activation system by tetracyclin has been developed, and this system may help the control of opiate secretion.We are constructing the cDNA including this system for making a new cell line.

近年来，微囊化细胞移植技术取得了进展。用于胶囊的半透膜允许低分子量物质的运输，但阻止免疫系统的体液和细胞成分的向内扩散。将前阿黑皮素基因转染的分泌β-内啡肽的Neuro 2A以5*10^6/ml的密度包封在聚合物胶囊中，并从雄性Sprague-Dawley大鼠的枕-寰交界处移植到脊髓CSF间隙中。三个测痛试验-夹尾试验、热板试验和电刺激试验-表明，移植后具有包封Neuro 2A的大鼠对疼痛的敏感性显著低于对照动物。形态学观察显示，微囊化细胞移植入脑脊液后可存活一个月。然而，日本猴CSF空间中的包封Neuro 2A显示出比大鼠差得多的活力。我们推测可能是补体或天然抗体穿透包膜攻击异种细胞。因此，将包封的致敏绵羊红细胞在10%人血清培养基中孵育。为了评估溶血的程度，测量介质波长（541 nm）处的吸光度。结果表明，由PM 30（Amicon公司;截留分子量为30 kD）和K6305（Kuraray公司;截留分子量为13 kD）组成的双层膜可抑制补体的穿透。接下来我们将尝试在日本猴的CSF间隙中移植含有Neuro 2A的双膜胶囊。在我们的细胞治疗中，控制阿片样物质的分泌量有点困难。然而，最近，四环素的转录激活系统已经被开发出来，该系统可能有助于控制阿片类物质的分泌，我们正在构建包含该系统的cDNA，以制备新的细胞系。