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新鲜与冷冻保存的猪中脑腹侧细胞移植于大鼠帕金森病模型中的比较。

Comparison of fresh and cryopreserved porcine ventral mesencephalon cells transplanted in A rat model of Parkinson's disease.

作者信息

Jacoby Douglas B, Lindberg Charles, Ratliff Judson, Wetzel Kristie, Stewart Gregory R, Dinsmore Jonathan

机构信息

Department of Cell Transplantation, Diacrin Inc., Charlestown, Massachusetts, USA.

出版信息

J Neurosci Res. 2002 Aug 1;69(3):382-96. doi: 10.1002/jnr.10297.

Abstract

To evaluate whether cryopreservation of porcine ventral mesencephalon cells influences graft survival and function in vivo, we have transplanted either freshly prepared or cryopreserved cells into the striatum of 6-hydroxydopamine-lesioned rats. A single cell suspension of porcine ventral mesencephalon cells from the same isolation either was stored at 4 degrees C and transplanted the next day or was cryopreserved for 4 weeks in liquid nitrogen vapor. The cryopreserved cells were then rapidly thawed, rinsed, and transplanted in the same manner as the fresh cells, with the same dose of viable cells. All animals received daily injections of cyclosporin A to prevent xenograft rejection. To monitor graft function, amphetamine-induced rotation was measured every 3 weeks between 6 and 15 weeks posttransplantation. After sacrifice at 15 weeks posttransplantation, histological methods were used to compare fresh cell and cryopreserved cell transplants with respect to graft survival, differentiation and integration, and host immune response. Cryopreserved cells were found to be either equivalent or in some cases superior to fresh cells with respect to rotational correction, graft survival, graft volume, numbers of graft-derived dopaminergic neurons, and host immune responses. In conclusion, the results indicate that it is feasible to cryopreserve porcine ventral mesencephalon cells for long-term storage of cells prior to transplantation in an animal model of Parkinson's disease.

摘要

为了评估猪中脑腹侧细胞的冷冻保存是否会影响其在体内移植后的存活及功能,我们将新鲜制备的或冷冻保存的细胞移植到6-羟基多巴胺损伤大鼠的纹状体中。来自同一分离物的猪中脑腹侧细胞单细胞悬液,一部分在4℃保存并于次日移植,另一部分在液氮蒸汽中冷冻保存4周。然后将冷冻保存的细胞快速解冻、冲洗,并以与新鲜细胞相同的方式、相同剂量的活细胞进行移植。所有动物每天注射环孢素A以防止异种移植排斥。为监测移植功能,在移植后6至15周期间每3周测量一次苯丙胺诱导的旋转行为。在移植后15周处死动物后,采用组织学方法比较新鲜细胞移植和冷冻保存细胞移植在移植存活、分化和整合以及宿主免疫反应方面的情况。结果发现,在旋转行为纠正、移植存活、移植体积、移植来源的多巴胺能神经元数量以及宿主免疫反应方面,冷冻保存的细胞与新鲜细胞相当,在某些情况下甚至更优。总之,结果表明在帕金森病动物模型中,移植前对猪中脑腹侧细胞进行冷冻保存以实现细胞的长期储存是可行的。

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