Butler P E, Lee W P, Sims C D, Randolph M A, Vacanti C A, Yaremchuk M J
Division of Plastic Surgery at Massachusetts General Hospital, Boston 02114, USA.
Plast Reconstr Surg. 1998 Jul;102(1):161-8; discussion 169-70. doi: 10.1097/00006534-199807000-00025.
A murine model of skeletal tissue transplantation was developed to study the allograft rejection process in mice for limb allograft transplantation. Muscle, bone, and skin have been shown to be strong antigenic stimuli in vascularized allograft models, and cells from these sources were used for transplantation. Using enzymatic digestion, keratinocytes, myocytes, and osteocytes were harvested from B10.A mice tissues, dissociated into single cells, and then grown in culture for 14 to 21 days. Each cell type was marked with an intracellular fluorescent marker before transplantation of the cells into pockets in the rectus abdominis muscle of a syngenic host. All cell types remained viable and were detectable 2 weeks following transplantation when examined histologically and observed under a fluorescent microscope. Transplanted osteocytes were found to produce bone 8 weeks following transplantation. These results demonstrate that individual cells transplanted into muscle pockets survive and have the ability to produce extracellular matrix in this mouse model of skeletal tissue transplantation. Use of this model will allow transplantation of the cellular components comprising limb allografts to study the relative antigenicities and the rejection of the separate cells with the advanced immunologic techniques available for mice. A better understanding of immunologic responses to these individual tissue components may enable specific donor tissue or host immune modification to achieve skeletal tissue transplantation without immunosuppression. These findings are particularly valuable to the field of tissue engineering where allogeneic cells may be used in cell/polymer constructs for reconstructive procedures.
为了研究小鼠肢体同种异体移植中的同种异体排斥过程，建立了一种骨骼组织移植的小鼠模型。在血管化同种异体移植模型中，肌肉、骨骼和皮肤已被证明是强烈的抗原刺激物，并且来自这些来源的细胞被用于移植。通过酶消化从B10.A小鼠组织中收获角质形成细胞、肌细胞和骨细胞，将其解离成单细胞，然后在培养中生长14至21天。在将细胞移植到同基因宿主的腹直肌囊袋之前，每种细胞类型都用细胞内荧光标记物进行标记。当进行组织学检查并在荧光显微镜下观察时，所有细胞类型在移植后2周仍保持存活且可检测到。移植后8周发现移植的骨细胞产生了骨组织。这些结果表明，在这个骨骼组织移植的小鼠模型中，移植到肌肉囊袋中的单个细胞能够存活并具有产生细胞外基质的能力。使用这个模型将允许移植构成肢体同种异体移植的细胞成分，以利用现有的先进小鼠免疫技术研究相对抗原性和单个细胞的排斥反应。更好地理解对这些单个组织成分的免疫反应可能使特定的供体组织或宿主免疫修饰能够在不进行免疫抑制的情况下实现骨骼组织移植。这些发现对于组织工程领域特别有价值，在该领域中，同种异体细胞可用于细胞/聚合物构建体以进行重建手术。
