Laboratory of Biochemistry and Cellular Engineering, Division of NanoBio Technology, Daegu Gyeongbuk Institute of Science and Technology, Daegu 711-873, Korea.
BMB Rep. 2013 May;46(5):276-81. doi: 10.5483/bmbrep.2013.46.5.196.
In this study, we aimed to compare the morphogenetic and neuronal characteristics between monolayer cells and spheroids. For this purpose, we established spheroid formation by growing SH-SY5Y cells on the hydrophobic surfaces of thermally-collapsed elastin-like polypeptide. After 4 days of culture, the relative proliferation of the cells within spheroids was approximately 92% of the values for monolayer cultures. As measured by quantitative assays for mRNA and protein expressions, the production of synaptophysin and neuronspecific enolase (NSE) as well as the contents of cell adhesion molecules (CAMs) and extracellular matrix (ECM) proteins are much higher in spheroids than in monolayer cells. Under the all-trans-retinoic acid (RA)-induced differentiation condition, spheroids extended neurites and further up-regulated the expression of synaptophysin, NSE, CAMs, and ECM proteins. Our data indicate that RA-differentiated SH-SY5Y neurospheroids are functionally matured neuronal architectures.
在这项研究中，我们旨在比较单层细胞和球体之间的形态发生和神经元特征。为此，我们通过在热塌陷弹性蛋白样多肽的疏水面上培养 SH-SY5Y 细胞来建立球体形成。培养 4 天后，球体中细胞的相对增殖率约为单层培养物的 92%。通过定量测定 mRNA 和蛋白质表达，发现突触小泡蛋白和神经元特异性烯醇化酶（NSE）的产生以及细胞黏附分子（CAM）和细胞外基质（ECM）蛋白的含量在球体中均高于单层细胞。在全反式视黄酸（RA）诱导分化条件下，球体延伸出神经突，并进一步上调突触小泡蛋白、NSE、CAM 和 ECM 蛋白的表达。我们的数据表明，RA 分化的 SH-SY5Y 神经球体是功能成熟的神经元结构。
