Y chromosome detection of three-dimensional tissue-engineered skeletal muscle constructs in a syngeneic rat animal model
Y chromosome detection of three-dimensional tissue-engineered skeletal muscle constructs in a syngeneic rat animal model
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DOI：
10.3727/000000004772664888
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发表时间：
2004-01-01
影响因子：
3.3
通讯作者：
Bach, AD
中科院分区：
文献类型：
--
作者：
Beier, JP;Kneser, U;Bach, AD
由于缺乏功能性天然组织替代，对因创伤或肿瘤消融而损失的肌肉组织进行手术重建受到限制。此外，到目前为止，大多数遗传性或后天性肌肉疾病都缺乏足够的治疗，因为只有很少的替代方案可以恢复丢失的肌肉组织的功能。对这些组织进行工程改造并移植到功能障碍部位可能是一种替代方法，并且可以替代此类受损或衰竭的骨骼肌组织。尝试重建一些人体组织和器官的技术（组织工程）最近已被引入临床实践。以往的移植研究面临的一个主要问题是整合后移植细胞的检测能力。在同系大鼠模型中使用 Y 染色体原位杂交技术可以原位移植细胞构建体，无需对细胞进行操作，不会产生排斥或免疫抑制，但提供了不可稀释的遗传标记来识别移植的细胞。我们研究的目的是通过移植在三维 (3D) 纤维蛋白基质中预培养的原代成肌细胞来在体内创建功能性骨骼肌组织，并使用 Y 染色体检测技术确定移植细胞的命运。 3D 成肌细胞培养物源自雄性供体大鼠，培养 7 天后，我们将 3D 细胞构建体原位移植到同系雌性大鼠股薄肌内产生的肌肉缺损中。抗结蛋白免疫染色和Y染色体原位杂交表明移植的雄性成肌细胞在雌性受体动物体内存活并整合，从而证明了这种方法在组织工程和细胞移植研究中的可行性。
Surgical reconstruction of muscle tissue lost by trauma or tumor ablation is limited by the lack of availability of functional native tissue substitution. Moreover, so far most inherited or acquired muscle diseases are lacking sufficient treatment, because only few alternatives exist to provide functional restoration of lost muscle tissues. Engineering those tissues and transplantation into sites of dysfunction may be an alternative approach and may allow replacement of such damaged or failing skeletal muscle tissues. Techniques attempting reconstruction of some human tissues and organs (tissue engineering) have been introduced into clinical practice recently. One major problem that previous transplantation studies were facing is the ability of detection of transplanted cells after integration. Using the Y chromosome in situ hybridization technique in a syngeneic rat model allows transplantation of cell constructs orthotopically, without manipulation of the cells, with no rejection or inummosuppression being implied, but providing a nondilutable genetic marker to identify transplanted cells. The purpose of our study was to create functional skeletal muscle tissue in vivo using the transplantation of primary myoblasts precultivated within a three-dimensional (3D) fibrin matrix and to determine the fate of the transplanted cells using the Y chromosome detection technique. 3D myoblast cultures were established derived from male donor rats and after 7 days of cultivation we performed an orthotopic transplantation of 3D cell constructs into a created muscle defect within the gracilis muscle of syngeneic female rats. Anti-desmin immunostaining and Y chromosome in situ hybridization indicated the survival and integration of transplanted male myoblasts into the female recipient animal, thus demonstrating the feasibility of this approach in tissue engineering and the research of cell transplantation in general.