Lentiviral Delivery of LMX1a Enhances Dopaminergic Phenotype in Differentiated Human Bone Marrow Mesenchymal Stem Cells
Lentiviral Delivery of LMX1a Enhances Dopaminergic Phenotype in Differentiated Human Bone Marrow Mesenchymal Stem Cells
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DOI：
10.1089/scd.2008.0138
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发表时间：
2009-05-01
影响因子：
4
通讯作者：
Offen, Daniel
中科院分区：
文献类型：
--
作者：
Barzilay, Ran;Ben-Zur, Tali;Offen, Daniel
人骨髓间充质干细胞(MSCs)存在于骨髓中，具有沿间充质谱系(脂肪、骨和软骨)分化的能力。最近的研究表明，这些细胞也有可能向神经外胚层分化。利用慢病毒基因传递，我们试图通过传递LMX1a来重新编程骨髓来源的MSCs向多巴胺能分化，据报道，LMX1a在发育动物模型和胚胎干细胞中都是多巴胺能分化的关键角色。携带荧光报告基因的细胞转导证实了基因传递的效率。转导LMX1a的细胞在分化培养液中孵育后，LMX1a蛋白集中在细胞核内，特异性多巴胺能发育基因表达上调。此外，与非转导细胞相比，转导细胞表达了更高水平的酪氨酸羟基酶，这是合成多巴胺的限速酶，并分泌了显著更高的多巴胺水平。因此，我们提出了一种新的策略，以促进骨髓来源的MSCs的多巴胺能分化，作为未来帕金森患者自体移植的可能细胞来源。
Human mesenchymal stem cells (MSCs) reside in the bone marrow and are known for their ability to differentiate along the mesenchymal lineage (fat, bone, and cartilage). Recent works have suggested the possibility that these cells are also capable of differentiating toward the neuroectodermal lineage. Using lentiviral gene delivery, we sought to reprogram the bone marrow-derived MSCs toward dopaminergic differentiation through delivery of LMX1a, which was reported to be a key player in dopaminergic differentiation in both developmental animal models and embryonic stem cells. Transduction of cells with fluorescent reporter genes confirmed efficiency of gene delivery. On incubation of the LMX1a transduced cells in differentiation medium, the LMX1a protein was concentrated in the cells' nuclei and specific dopaminergic developmental genes were upregulated. Moreover, the transduced cells expressed higher levels of tyrosine hydroxylase, the rate limiting enzyme in dopamine synthesis, and secreted significantly higher level of dopamine in comparison to nontransduced cells. We hereby present a novel strategy to facilitate the dopaminergic differentiation of bone marrow-derived MSCs as a possible cell source for autologous transplantation for Parkinsonian patients in the future.