Stanko Peter, Kaiserova Katarina, Altanerova Veronika, Altaner Cestmir
Department of Stomatology and Maxillofacial Surgery, Faculty of Medicine, Comenius University in Bratislava, Bratislava, Slovak Republic.
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2014 Sep;158(3):373-7. doi: 10.5507/bp.2013.078. Epub 2013 Oct 18.
Our aims were to characterize human mesenchymal stem cells isolated from various tissues by pluripotent stem cells gene expression profile.
Four strains of dental pulp stem cells (DP-MSCs) were isolated from dental pulp tissue fragments adhered to plastic tissue culture dishes. Mesenchymal stem cells derived from umbilical cord tissue (UBC-MSCs) were isolated with the same technique. Bone marrow derived mesenchymal stem cells (BM-MSCs) were isolated from nucleated cells of bone marrow obtained by density gradient centrifugation. Human mesenchymal stem cells from adipose tissue (AT-MSCs) were isolated by collagenase digestion. All kinds of MSCs used in this study were cultivated in low glucose DMEM containing 5% or human platelet extract. All stem cell manipulation was performed in GMP conditions. Expression of 15 pluripotent stem cells genes on the level of proteins was measured by Proteome Profiler Human Pluripotent Stem Cell Array. Induction of MSCs to in vitro differentiation to adipocytes, osteoblasts, chondroblasts was achieved by cultivation of cells in appropriate differentiation medium.
All MSCs tested were phenotypically similar and of fibroblastoid morphology. DP-MSCs and UBC-MSCs were more proliferative than bone marrow BM-MSCs and AT-MSCs. Protein expression of 15 genes typical for pluripotent stem cells distinguished them into two groups. While the gene expression profiles of BM-MSC, AT-MSCs and UBC-MSCs were similar, DP-MSCS differed in relative gene expression on the level of their products in several genes.
Dental pulp mesenchymal stem cells cultivated in vitro under the same conditions as MSCs from bone marrow, adipose tissue and umbilical cord tissue can be distinguished by pluripotent stem cell gene expression profile.
我们的目的是通过多能干细胞基因表达谱来表征从各种组织中分离出的人间充质干细胞。
从粘附于塑料组织培养皿的牙髓组织碎片中分离出四株牙髓干细胞(DP-MSCs)。采用相同技术分离出源自脐带组织的间充质干细胞(UBC-MSCs)。从通过密度梯度离心获得的骨髓有核细胞中分离出骨髓来源的间充质干细胞(BM-MSCs)。通过胶原酶消化分离出人脂肪组织间充质干细胞(AT-MSCs)。本研究中使用的所有种类的间充质干细胞均在含有5%人血小板提取物的低糖DMEM中培养。所有干细胞操作均在GMP条件下进行。通过蛋白质组分析人多能干细胞阵列检测15种多能干细胞基因在蛋白质水平上的表达。通过在适当的分化培养基中培养细胞,诱导间充质干细胞体外分化为脂肪细胞、成骨细胞、成软骨细胞。
所有测试的间充质干细胞在表型上相似,呈成纤维细胞样形态。DP-MSCs和UBC-MSCs比骨髓BM-MSCs和AT-MSCs增殖能力更强。15种多能干细胞典型基因的蛋白质表达将它们分为两组。虽然BM-MSC、AT-MSCs和UBC-MSCs的基因表达谱相似,但DP-MSCS在几个基因的产物水平上的相对基因表达有所不同。
在与骨髓、脂肪组织和脐带组织的间充质干细胞相同条件下体外培养的牙髓间充质干细胞可以通过多能干细胞基因表达谱来区分。