Habisch H-J, Fiedler J, Ludolph A C, Storch A, Brenner R E
Department of Neurology, University of Ulm, Ulm, Germany.
Cytotherapy. 2008;10(8):824-33. doi: 10.1080/14653240802474331.
Bone marrow (BM)-derived mesenchymal stromal cells (MSC) are promising candidate cells for the development of neuroregenerative therapies. We have previously introduced the pro-neural conversion of human MSC to neural stem cell-like cells (m-NSC) by culturing them in suspension culture under serum-free conditions.
In the present study, we used a modified Boyden chamber assay to study the influence of various chemoattractants and extracellular matrix components on MSC and m-NSC migration in vitro. The underlying mechanisms were investigated further by applying real-time reverse transcriptase (RT)-polymerase chain reaction (PCR) and flow cytometry.
The basal migration of m-NSC was significantly reduced compared with MSC (six versus 27 out of 10,000 cells migrated within 6 h). We evaluated the effects of bone morphogenic protein 2 (BMP2), insulin-like growth factor 1 (IGF1), platelet-derived growth factor bb (PDGFbb), vascular endothelial growth factor (VEGFa), and stromal cell-derived factor 1 (SDF1) on the migration potential of both cell types and PDGFbb proved to be the most potent stimulant of migration (235 versus 198 m-NSC or MSC migrated). Adhesion of m-NSC to the filter membrane was delayed and not affected by IGF1 or PDGFbb: 90% of MSC, but only 20% of m-NSC, adhered within 1 h, with 90% of m-NSC adhering within 3 h. However, real-time RT-PCR and flow cytometry revealed an up-regulation of the PDGF receptor B following conversion. Coating the membranes with collagen type I or hyaluronan also significantly influenced cell migration.
We could identify major chemoattractive factors for m-NSC and gained partial insight into the complex processes involved in migration of neurally converted cells.
骨髓（BM）来源的间充质基质细胞（MSC）是神经再生治疗开发中很有前景的候选细胞。我们之前通过在无血清条件下进行悬浮培养，将人MSC促神经转化为神经干细胞样细胞（m-NSC）。
在本研究中，我们使用改良的Boyden小室试验来研究各种趋化因子和细胞外基质成分对体外MSC和m-NSC迁移的影响。通过实时逆转录（RT）-聚合酶链反应（PCR）和流式细胞术进一步研究其潜在机制。
与MSC相比，m-NSC的基础迁移显著降低（6小时内每10000个细胞中迁移的细胞数分别为6个和27个）。我们评估了骨形态发生蛋白2（BMP2）、胰岛素样生长因子1（IGF1）、血小板衍生生长因子bb（PDGFbb）、血管内皮生长因子（VEGFa）和基质细胞衍生因子1（SDF1）对两种细胞类型迁移潜能的影响，结果证明PDGFbb是最有效的迁移刺激物（迁移的m-NSC或MSC分别为235个和198个）。m-NSC与滤膜的黏附延迟，且不受IGF1或PDGFbb影响：90%的MSC在1小时内黏附，而m-NSC只有20%在1小时内黏附，90%的m-NSC在3小时内黏附。然而，实时RT-PCR和流式细胞术显示转化后血小板衍生生长因子受体B上调。用I型胶原或透明质酸包被滤膜也显著影响细胞迁移。
我们能够确定m-NSC的主要趋化因子，并对神经转化细胞迁移所涉及的复杂过程有了部分了解。
