脂肪基质细胞与颅缝衍生间充质细胞的旁分泌相互作用。

Paracrine interaction between adipose-derived stromal cells and cranial suture-derived mesenchymal cells.

机构信息

Stanford, Calif. From the Hagey Laboratory for Pediatric Regenerative Medicine, Department of Surgery, Plastic and Reconstructive Surgery Division, Stanford University School of Medicine.

出版信息

Plast Reconstr Surg. 2010 Sep;126(3):806-821. doi: 10.1097/PRS.0b013e3181e5f81a.

Abstract

BACKGROUND

Adipose-derived stromal cells are a potential cell source for the successful healing of skeletal defects. In this study, the authors sought to investigate the potential for cranial suture-derived mesenchymal cells to promote the osteogenic differentiation of adipose-derived stromal cells. Various reports have previously examined the unique in vitro attributes of suture-derived mesenchymal cells; this study sought to extend those findings.

METHODS

Suture-derived mesenchymal cells were isolated from wild-type mice (n = 30) from both fusing posterofrontal and patent sagittal sutures. Cells were placed in Transwell inserts with human adipose-derived stromal cells (n = 5 patients) with osteogenic differentiation medium with or without recombinant Noggin (10 to 400 ng/ml). Specific gene expression of osteogenic markers and Hedgehog pathway were assayed; standard osteogenic assays (alkaline phosphatase and alizarin red staining) were performed. All assays were performed in triplicate.

RESULTS

Both posterofrontal and sagittal suture-derived mesenchymal cells induced osteogenic differentiation of adipose-derived stromal cells (p < 0.05). Posterofrontal suture-derived mesenchymal cells induced adipose-derived stromal cell osteogenesis to a greater degree than sagittal suture-derived mesenchymal cells (p < 0.05). This was accompanied by an increase in bone morphogenetic protein expression (p < 0.05). Finally, recombinant Noggin mitigated the pro-osteogenic effects of co-culture accompanied by a reduction in Hedgehog signaling (p < 0.05).

CONCLUSIONS

Suture-derived mesenchymal cells secrete paracrine factors that induce osteogenic differentiation of multipotent stromal cells (human adipose-derived stromal cells). Cells derived from the fusing posterofrontal suture do this to a significantly greater degree than cells from the patent sagittal suture. Enhanced bone morphogenetic protein and Hedgehog signaling may underlie this paracrine effect.

摘要

背景

脂肪来源的基质细胞是成功修复骨缺损的潜在细胞来源。在这项研究中,作者试图研究颅缝衍生间充质细胞促进脂肪来源的基质细胞成骨分化的潜力。先前有许多报道研究了缝间充质细胞的独特体外特性;本研究旨在扩展这些发现。

方法

从融合额眶缝和开放矢状缝的野生型小鼠(n=30)中分离出颅缝衍生间充质细胞。将细胞置于 Transwell 插入物中,与接受成骨分化培养基的人脂肪来源的基质细胞(n=5 名患者)共培养,其中含有或不含重组 Noggin(10 至 400ng/ml)。检测成骨标志物和 Hedgehog 通路的特定基因表达;进行标准成骨测定(碱性磷酸酶和茜素红染色)。所有测定均重复三次。

结果

额眶缝和矢状缝衍生间充质细胞均诱导脂肪来源的基质细胞成骨分化(p<0.05)。额眶缝衍生间充质细胞诱导脂肪来源的基质细胞成骨作用强于矢状缝衍生间充质细胞(p<0.05)。这伴随着骨形态发生蛋白表达的增加(p<0.05)。最后,重组 Noggin 减轻了共培养的促成骨作用,同时降低了 Hedgehog 信号(p<0.05)。

结论

颅缝衍生间充质细胞分泌旁分泌因子,诱导多能基质细胞(人脂肪来源的基质细胞)成骨分化。来自融合的额眶缝的细胞比来自开放矢状缝的细胞具有更大的程度。增强的骨形态发生蛋白和 Hedgehog 信号可能是这种旁分泌作用的基础。

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