Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):2924-2934. doi: 10.26355/eurrev_201904_17572.
To evaluate the different behavior of two different human adult adipocytes derived stem cells (hASCs) during proliferation and osteogenic differentiation.
Human adult adipocytes stem cells (hAT-SCs) from visceral (hAV-SCs) and subcutaneous (hAS-SCs) sites were obtained after surgery procedures of seven patients. All samples were fully investigated and the different proliferation rates were evaluated. All MSCs clusters were cultured with an osteogenic and adipogenic differentiation medium. Homogeneous pools of Mesenchymal Stem Cells (MSCs) were confirmed by Flow-Cytometry Analysis (FACS) and Spectrophotometric Assay. The differentiated cells were eventually assessed for the expression of Alkaline Phosphatase (ALP), Alizarin Red (AR) and Oil Red-O (OR-O) detection, and analyzed by the Spectrophotometric Assay. After osteogenic differentiation, the cell clusters were incubated and analyzed with Real Time-Polymerase Chain Reaction (qRT-PCR) and fluorescence microscopy.
The FACS analysis performed on hAT-SCs confirmed the homogenous presence of MSCs in all samples. The ALP, AR stain confirmed the osteogenic differentiation capacity of MSCs towards osteoblast-like-cells. The colorimetric cell metabolic activity (MTS) assay showed an increase in the proliferation rate with different values in both sets hAS-SCs vs. hAV-SCs.
These in vitro findings of both hAS-SCs and hAV-SCs suggested an important role of these stem cells for future clinical use in bone regeneration. Indeed, the final outcomes suggested a better performance of cells coming from subcutaneous adipose tissue vs. those from visceral fat tissue.
评估两种不同的人类成体脂肪来源干细胞（hASCs）在增殖和成骨分化过程中的不同行为。
从 7 名患者的手术部位获得了内脏（hAV-SCs）和皮下（hAS-SCs）来源的人成体脂肪干细胞（hAT-SCs）。对所有样本进行了全面研究，并评估了不同的增殖率。所有 MSC 集落均用成骨和成脂分化培养基培养。通过流式细胞术分析（FACS）和分光光度法确认同质的间充质干细胞（MSCs）池。分化细胞最终通过碱性磷酸酶（ALP）、茜素红（AR）和油红-O（OR-O）检测进行评估，并通过分光光度法进行分析。成骨分化后，用实时聚合酶链反应（qRT-PCR）和荧光显微镜孵育和分析细胞集落。
对 hAT-SCs 进行的 FACS 分析证实了所有样本中 MSC 的同质存在。ALP、AR 染色证实了 MSC 向成骨样细胞的成骨分化能力。比色细胞代谢活性（MTS）测定显示增殖率增加，两组 hAS-SCs 与 hAV-SCs 的值不同。
这些 hAS-SCs 和 hAV-SCs 的体外发现表明这些干细胞在未来的骨再生临床应用中具有重要作用。事实上，最终结果表明，来自皮下脂肪组织的细胞比来自内脏脂肪组织的细胞表现更好。
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