Henriksson Helena B, Svanvik Teresia, Jonsson Marianne, Hagman Margret, Horn Michael, Lindahl Anders, Brisby Helena
The Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Spine (Phila Pa 1976). 2009 Jan 15;34(2):141-8. doi: 10.1097/BRS.0b013e31818f8c20.
Experimental and descriptive study of a xenotransplantation model in minipigs.
To study survival and function of human mesenchymal stem cells (hMSCs) after transplantation into injured porcine spinal discs, as a model for cell therapy.
Biologic treatment options of the intervertebral disc are suggested for patients with chronic low back pain caused by disc degeneration.
Three lumbar discs in each of 9 minipigs were injured by aspiration of the nucleus pulposus (NP), 2 weeks later hMSCs were injected in F12 media suspension (cell/med) or with a hydrogel carrier (Puramatrix) (cell/gel). The animals were sacrificed after 1, 3, or 6 months. Disc appearance was visualized by magnetic resonance imaging. Immunohistochemistry methods were used to detect hMSCs by antihuman nuclear antibody staining, and further performed for Collagen II, Aggrecan, and Collagen I. SOX 9, Aggrecan, Versican, Collagen IA, and Collagen IIA and Collagen IIB human mRNA expression was analyzed by real-time PCR.
At magnetic resonance imaging all injured discs demonstrated degenerative signs. Cell/gel discs showed fewer changes compared with cell/med discs and only injured discs at later time points. hMSCs were detected in 9 of 10 of the cell/gel discs and in 8 of 9 of the cell/med discs. Immunostaining for Aggrecan and Collagen type II expression were observed in NP after 3 and 6 months in gel/cell discs and colocalized with the antihuman nuclear antibody. mRNA expression of Collagen IIA, Collagen IIB, Versican, Collagen 1A, Aggrecan, and SOX9 were detected in both cell/med and cell/gel discs at the time points 3 and 6 months by real-time PCR.
hMSCs survive in the porcine disc for at least 6 months and express typical chondrocyte markers suggesting differentiation toward disc-like cells. As in autologous animal models the combination with a three-dimensional-hydrogel carrier seems to facilitate differentiation and survival of MSCs in the disc. Xenotransplantation seems to be valuable in evaluating the possibility for human cell therapy treatment for intervertebral discs.
小型猪异种移植模型的实验性和描述性研究。
研究人骨髓间充质干细胞（hMSCs）移植到损伤的猪椎间盘后其存活及功能情况，以此作为细胞治疗的模型。
对于因椎间盘退变导致慢性下腰痛的患者，建议采用椎间盘的生物治疗方法。
对9只小型猪，每只猪的3个腰椎间盘通过抽吸髓核进行损伤，2周后将hMSCs以F12培养基悬浮液（细胞/培养基）或与水凝胶载体（Puramatrix）（细胞/凝胶）混合的方式注入。在1、3或6个月后处死动物。通过磁共振成像观察椎间盘外观。采用免疫组织化学方法通过抗人核抗体染色检测hMSCs，并进一步检测Ⅱ型胶原、聚集蛋白聚糖和Ⅰ型胶原。通过实时聚合酶链反应分析SOX 9、聚集蛋白聚糖、多功能蛋白聚糖、Ⅰ型胶原A、Ⅱ型胶原A和Ⅱ型胶原B的人信使核糖核酸表达。
在磁共振成像中，所有损伤的椎间盘均显示退变迹象。与细胞/培养基组椎间盘相比，细胞/凝胶组椎间盘变化较少，且仅在后期时间点的损伤椎间盘中出现变化。在10个细胞/凝胶组椎间盘中的9个以及9个细胞/培养基组椎间盘中的8个检测到了hMSCs。在凝胶/细胞组椎间盘中，3个月和6个月后在髓核中观察到聚集蛋白聚糖和Ⅱ型胶原表达的免疫染色，且与抗人核抗体共定位。通过实时聚合酶链反应在3个月和6个月时间点在细胞/培养基组和细胞/凝胶组椎间盘中均检测到Ⅱ型胶原A、Ⅱ型胶原B、多功能蛋白聚糖、Ⅰ型胶原A、聚集蛋白聚糖和SOX9的信使核糖核酸表达。
hMSCs在猪椎间盘中存活至少6个月，并表达典型的软骨细胞标志物，提示其向椎间盘样细胞分化。与自体动物模型一样，与三维水凝胶载体联合似乎有助于间充质干细胞在椎间盘中的分化和存活。异种移植在评估人类椎间盘细胞治疗可能性方面似乎具有重要价值。
