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Comparative Analysis of Wharton's Jelly Mesenchymal Stem Cell (WJ-MSCs) Isolated Using Explant and Enzymatic Methods

机译:沃顿果冻间充质干细胞(WJ-MSCs)的比较分析使用外植体和酶法分离

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Wharton's Jelly is one of the best sources for mesenchymal stem cells. Human Wharton's Jelly Mesenchymal Stem Cells (hWJ-MSCs) have high proliferation, multi-lineage differentiation potential, and do not produce any teratogen or carcinogen. These characteristics make hWJ-MSCs become suitable for regenerative medicine. Some methods were developed to isolate hWJ-MSCs from umbilical cord, such as explant method and enzymatic method. This study aims to characterize hWJ-MSCs which are isolated by two different methods, explant attachment method and enzymatic method. hWJ-MSCs isolation was performed through explant method and enzymatic method using trypsin, hyaluronidase and collagenase type 1 with certain ratio of concentration. Isolated hWJ-MSCs was characterized using flow cytometer to detect the expression of CD44, CD90, CD105, CD73 and negative lineage. MSCs differentiation assay was performed to analyze adipogenic, chondrogenic and osteogenic cells lineage. We successfully isolated hWJ-MSCs from umbilical cord through enzymatic and explant methods. Immunophenotyping assay through flow cytometry analysis showed high purity of WJ-MSCs. The isolated hWJ-MSCs from both methods showed positive expression of CD44, CD90, CD105, and CD73. The isolated hWJ-MSCs exhibited capacity to differentiate into adipocyte, chondrocyte, and osteocyte cells. hWJ-MSCs isolated through explant and enzymatic method have high proliferation capacity and be able to differentiate into three different lineage cells. Both methods explant attachment and enzymatic methods are efficiently produced hWJ-MSCs.
机译:沃顿果冻是间充质干细胞的最佳来源之一。人沃顿的果冻间充质干细胞(HWJ-MSCs)具有高增殖,多谱分化潜力,并且不会产生任何致畸原或致癌物质。这些特性使HWJ-MSCs适用于再生医学。开发了一些方法,用于从脐带中分离HWJ-MSC,例如外植体方法和酶法。本研究旨在表征由两种不同方法分离的HWJ-MSC,外显性附着法和酶法。通过使用胰蛋白酶,透明质酸酶和胶原蛋白酶1的外胰岛方法和酶法进行HWJ-MSCS分离,具有一定的浓度比例。孤立的HWJ-MSCs使用流式细胞仪进行了分析,以检测CD44,CD90,CD105,CD73和负谱系的表达。进行MSCS分化测定以分析脂肪发生,软骨内和骨质发生细胞谱系。我们通过酶促和外盐方法成功地将HWJ-MSC从脐带上分离出来。通过流式细胞术分析的免疫表型测定显示出高纯度的WJ-MSC。来自两种方法的分离的HWJ-MSC显示CD44,CD90,CD105和CD73的阳性表达。分离的HWJ-MSCs表现出分化成脂肪细胞,软骨细胞和骨细胞细胞的能力。通过外植体和酶法分离的HWJ-MSC具有高增殖能力,能够分为三种不同的谱系细胞。两种方法都是有效生产HWJ-MSCs的脱钙附着和酶法。

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