Laboratory of Experimental Urology, Department of Development and Regeneration, University of Leuven Leuven, Belgium ; Laboratory of Experimental Gynecology, Department of Development and Regeneration, University of Leuven Leuven, Belgium.
Laboratory of Experimental Urology, Department of Development and Regeneration, University of Leuven Leuven, Belgium.
Sex Med. 2013 Aug;1(1):3-15. doi: 10.1002/sm2.1.
Adipose tissue-derived stem cells (ADSCs) herald tremendous promise for clinical application in a wide range of injuries and diseases. Several preclinical reports demonstrate their efficacy in the treatment of cavernous nerve (CN) injury-induced erectile dysfunction in rats. It was recently illustrated that these effects were established as a result of ADSC migration to the major pelvic ganglion (MPG) where these cells induced neuroregeneration in loco.
The study aims to identify chemotactic factors in the MPG following injury and to match upregulated chemokines to their respective receptors in human ADSC on the genomic, structural, and functional levels.
Quantitative real-time polymerase chain reaction, fluorescence-activated cell sorting (FACS), intracellular FACS, immunofluorescence microscopy, migration assays, and calcium imaging were used in this study.
The main outcomes are chemokine expression in the MPG following CN injury, and the functional and structural presence of chemokine receptors in ADSC.
CCR4, CX3CR1, and XCR1 are functionally and structurally present in human ADSC, and are activated by the chemokines CCL2, CX3CL1, and XCL1 respectively, which are upregulated in the MPG following CN injury. CXCR4 and its ligand CXCL12 (SDF1) are likely no major homing factors for ADSC. Expression of chemokine receptor mRNA in ADSC did not necessarily translate into receptor presence at the cell surface and/or functional activation of these receptors. Most of the expressed chemokine receptors were detected in the intracellular compartment of these cells.
We identified the ligand/chemokine receptor pairs CCL2/CCR4, CX3CL1/CX3CR1, and XCL1/XCR1 as potentially responsible for ADSC homing toward the MPG following CN injury. The intracellular localization of various chemokine receptors likely indicates redirecting of chemokine receptors to the cell surface under specific cellular conditions. Furthermore, modification of expression of these receptors at the genomic level may potentially lead to improved migration toward injury sites and thus enhancement of treatment efficacy. Albersen M, Berkers J, Dekoninck P, Deprest J, Lue TF, Hedlund P, Lin C-S, Bivalacqua TJ, Van Poppel H, De Ridder D, and Van der Aa F. Expression of a distinct set of chemokine receptors in adipose tissue-derived stem cells is responsible for in vitro migration toward chemokines appearing in the major pelvic ganglion following cavernous nerve injury. Sex Med 2013;1:3-15.
脂肪组织来源的干细胞（ADSCs）在治疗广泛的损伤和疾病方面具有巨大的临床应用潜力。一些临床前报告表明，它们在治疗大鼠海绵体神经（CN）损伤引起的勃起功能障碍方面具有疗效。最近的研究表明，这些效果是由于 ADSC 迁移到主要盆神经节（MPG）而建立的，这些细胞在局部诱导了神经再生。
本研究旨在鉴定损伤后 MPG 中的趋化因子，并在基因组、结构和功能水平上将上调的趋化因子与其在人 ADSC 中的相应受体相匹配。
本研究采用定量实时聚合酶链反应、荧光激活细胞分选（FACS）、细胞内 FACS、免疫荧光显微镜、迁移实验和钙成像。
主要观察指标是 CN 损伤后 MPG 中趋化因子的表达，以及 ADSC 中趋化因子受体的功能和结构存在。
CCR4、CX3CR1 和 XCR1 在人 ADSC 中具有功能和结构，分别被趋化因子 CCL2、CX3CL1 和 XCL1 激活，这些趋化因子在 CN 损伤后在 MPG 中上调。CXCR4 及其配体 CXCL12（SDF1）可能不是 ADSC 的主要归巢因子。ADSC 中趋化因子受体 mRNA 的表达不一定转化为细胞表面的受体存在和/或这些受体的功能激活。这些细胞的大多数表达的趋化因子受体都存在于细胞内区室中。
我们确定了配体/趋化因子受体对 CCL2/CCR4、CX3CL1/CX3CR1 和 XCL1/XCR1，作为 ADSC 归巢到 CN 损伤后 MPG 的潜在原因。各种趋化因子受体的细胞内定位可能表明，在特定的细胞条件下，将趋化因子受体重新定向到细胞表面。此外，在基因组水平上修饰这些受体的表达可能潜在地导致向损伤部位的迁移增加，从而增强治疗效果。Albersen M、Berkers J、Dekoninck P、Deprest J、Lue TF、Hedlund P、Lin C-S、Bivalacqua TJ、Van Poppel H、De Ridder D 和 Van der Aa F. 在脂肪组织来源的干细胞中表达的一组特定的趋化因子受体负责体外向趋化因子迁移，这些趋化因子在海绵体神经损伤后出现在主要盆神经节中。性医学 2013；1:3-15。
