Baboon mesenchymal stem cells can be genetically modified to secrete human erythropoietin in vivo
Baboon mesenchymal stem cells can be genetically modified to secrete human erythropoietin in vivo
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DOI：
10.1089/10430340152480258
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发表时间：
2001-08-01
影响因子：
4.2
通讯作者：
Devine, SM
中科院分区：
文献类型：
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作者：
Bartholomew, A;Patil, S;Devine, SM
人骨髓间充质干细胞（MSCs）具有分化为软骨细胞、骨细胞、脂肪细胞和骨髓基质细胞等多种间充质细胞的能力。使用非人灵长类动物模型，我们评估非人灵长类动物间充质干细胞作为基因治疗的目标。狒狒骨髓间充质干细胞（bMSCs）培养骨髓穿刺液出现作为一个均匀的人口梭形细胞。骨髓间充质干细胞在体外可分化为脂肪细胞和骨细胞，在体内可分化为软骨细胞。用编码人促红细胞生成素（hEPO）基因和绿色荧光蛋白（GFP）基因的双顺反子载体对bMSC进行遗传修饰。骨髓间充质干细胞与逆转录病毒上清液孵育后，转导效率范围为72%至99%。植入前，体外每24小时每10（6）个细胞产生1.83 × 10（5）至7.12 × 10（5）mIU的hEPO。为了确定bMSC在体内表达hEPO的能力，将转导的bMSC肌内注射到NOD/SCID小鼠中。在一个单独的实验中，将转导的bMSCs加载到免疫隔离装置（IID）中，并通过手术植入自体或同种异体狒狒受体中。在NOD/SCID小鼠的血清中检测到人EPO长达28天，在5只狒狒的血清中检测到人EPO长达9至137天。NOD/SCID小鼠在肌内注射hEPO转导的bMSCs后红细胞比容急剧上升。表达hEPO达137天的狒狒经历了其血细胞比容的统计学显著（p < 0.04）升高。这些数据表明，非人灵长类动物MSC可以被工程化以递送分泌的和生物活性的基因产物。因此，人MSC可能是未来人类基因治疗试验的有效靶点。
Human mesenchymal stem cells (MSCs) are capable of differentiating into multiple mesenchymal lineages including chondrocytes, osteocytes, adipocytes, and marrow stromal cells. Using a nonhuman primate model, we evaluated nonhuman primate MSCs as targets for gene therapy. Baboon MSCs (bMSCs) cultured from bone marrow aspirates appeared as a homogeneous population of spindle-shaped cells. bMSCs were capable of differentiating into adipocytes and osteocytes in vitro and chondrocytes in vivo. bMSCs were genetically modified with a bicistronic vector encoding the human erythropoietin (hEPO) gene and the green fluorescent protein (GFP) gene. Transduction efficiencies ranged from 72 to 99% after incubation of MSCs with retroviral supernatant. Transduced cells produced from 1.83 X 10(5) to 7.12 X 10(5) mIU of hEPO per 10(6) cells per 24 hr in vitro before implantation. To determine the capacity of bMSCs to express hEPO in vivo, transduced bMSCs were injected intramuscularly in NOD/SCID mice. In a separate experiment, transduced bMSCs were loaded into immunoisolatory devices (IIDs) and surgically implanted into either autologous or allogeneic baboon recipients. Human EPO was detected in the serum of NOD/SCID mice for up to 28 days and in the serum of five baboons for between 9 and 137 days. NOD/SCID mice experienced sharp rises in hematocrit after intramuscular injection of hEPO-transduced bMSCs. The baboon that expressed hEPO for 137 days experienced a statistically significant (p < 0.04) rise in its hematocrit. These data demonstrate that nonhuman primate MSCs can be engineered to deliver a secreted and biologically active gene product. Therefore, human MSCs may be an effective target for future human gene therapy trials.