糖胺聚糖含量和II型胶原定位在l -抗坏血酸2-磷酸诱导脂肪源间充质干细胞成软骨分化中的作用

IF 0.5 Q4 MULTIDISCIPLINARY SCIENCES

Journal of Mathematical and Fundamental Sciences Pub Date : 2020-04-30 DOI:10.5614/j.math.fund.sci.2020.52.1.7

A. Barlian, N. L. W. E. Yanti

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引用次数: 2

摘要

已知l -抗坏血酸2-磷酸(LAA)可诱导软骨细胞分化。本研究的目的是通过分析糖胺聚糖(GAG)含量和II型胶原(Coll2)定位来分析LAA在脂肪源性间充质干细胞(ADSC)软骨分化中的作用。采用流式细胞术对ADSC进行表征，并在含有不同浓度LAA(0、25、50、100 μg/mL)的培养基中培养2、3、4周。用共聚焦显微镜免疫细胞化学(ICC)分析Coll2的定位。阿利新蓝染色定量GAG，茜素红S染色定量钙沉积。结果显示，ADSC细胞间充质干细胞(MSC)标记物阳性。Coll2定位于细胞质中，并随着LAA浓度的增加而增加。在100 μg/mL的LAA中，Coll2的定位强度最高。LAA诱导培养基中的ADSC中GAG含量高于对照组(LAA 0 μg/mL) (p < 0.05)。培养4周后，LAA浓度为25 μg/mL时钙沉积量最高(p < 0.05)，浓度越高钙沉积量越少。综上所述，LAA 100 μg/mL被认为是软骨分化的最佳LAA浓度。

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Glycosaminoglycans Content and Type II Collagen Localization in Chondrogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells Induced by L-Ascorbic Acid 2-Phosphate

L-ascorbic acid 2-phosphate (LAA) is known to induce chondrocyte differentiation. The objective of this study was to analyze the potency of LAA in chondrogenic differentiation of adipose-derived mesenchymal stem cells (ADSC) by analyzing the glycosaminoglycans (GAG) content and type II collagen (Coll2) localization. ADSC was characterized using flow cytometry and cultured in media containing various concentrations of LAA (0, 25, 50, 100 μg/mL) for 2, 3 and 4 weeks. Coll2 localization was analyzed by immunocytochemistry (ICC) using a confocal microscope. The quantification of GAG was performed by Alcian Blue staining and calcium deposition by Alizarin Red S staining. The results showed that ADSC was positive for mesenchymal stem cell (MSC) markers. Coll2 was localized in the cytoplasm and showed increasing abundance along with the increase of the LAA concentration. The highest intensity of Coll2 localization was shown in LAA 100 μg/mL. ADSC in LAA induction medium showed higher GAG content compared to the control group (LAA 0 μg/mL) (p < 0.05). The highest calcium deposit was shown by LAA 25 μg/mL after 4 weeks of culture (p < 0.05) and it decreased at higher concentrations. In conclusion, LAA 100 μg/mL is considered the optimum LAA concentration for chondrogenic differentiation.

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来源期刊

Journal of Mathematical and Fundamental Sciences MULTIDISCIPLINARY SCIENCES-

CiteScore

1.30

自引率

0.00%

发文量

审稿时长

24 weeks

期刊介绍： Journal of Mathematical and Fundamental Sciences welcomes full research articles in the area of Mathematics and Natural Sciences from the following subject areas: Astronomy, Chemistry, Earth Sciences (Geodesy, Geology, Geophysics, Oceanography, Meteorology), Life Sciences (Agriculture, Biochemistry, Biology, Health Sciences, Medical Sciences, Pharmacy), Mathematics, Physics, and Statistics. New submissions of mathematics articles starting in January 2020 are required to focus on applied mathematics with real relevance to the field of natural sciences. Authors are invited to submit articles that have not been published previously and are not under consideration elsewhere.