探討3T3纖維母細胞在三維微環境中的形狀

Analyzing the morphology of 3T3 fibroblasts in microenvironment

洪瑋嶸(Wei-Hong Hong)

指導教授 ： 陳義裕

國立臺灣大學/理學院/物理研究所/碩士(2010年)

https://doi.org/10.6342/NTU.2010.02419

摘要

我們用良好排列大小均一的孔洞製造了三維的明膠培養鷹架，並且在其內養著3T3纖維母細胞。我們對微環境如何影響細胞的形狀非常有興趣，所以我們用有螢光的phalloidin去染細胞內部的肌動蛋白，並且用共軛焦顯微術去取得我們要的細胞影像，為了看清楚我們的細胞，我們經由先改良原本的凝膠化學反應過程來降低我們鷹架本身強烈的自發螢光。另外我們做了一系列不同實驗來看微環境對細胞形狀的影響，如細胞養在二維的硬表面上、二維的明膠軟表面上、三維的膠原蛋白凝膠內，跟我們製造出的不同大小孔洞的培養鷹架內。我們將細胞的形狀分成三類，並且用橢球去近似細胞的形狀來量測細胞在不同維環境伸展的長度。從細胞伸展長度的趨勢來看，細胞在比較大的孔洞鷹架內，比較接近養在二維的明膠軟表面，這樣的結果暗示著細胞養在我們有曲率的鷹架內，他的行為是二維到三維培養的過渡階段。

關鍵字

三維培養 ； 3T3纖維母細胞 ； 形態學 ； 自發螢光 ； 明膠培養鷹架 ； 均一大小孔洞

並列摘要

We have created 3D ordered gelatin scaffolds with monodisperse pores and cultured 3T3 fibroblasts inside. We are interested to explore the effect of microenvironment on the cell morphologies. The morphologies of cells are observed through labeling the F-actin inside the cells with fluorescent phalloidin and imaged by a confocal microscope. To segment the cell properly, we first reduced the autofluorescence background from the scaffold by changing the crosslinking chemistry. We compared the cell morphologies in the following microenvironment - on a 2D hard surface, on a 2D soft gelatin surface, in a 3D collagen gel, and scaffolds of different pore sizes. We found that the cells exhibit wide range of morphologies in different microenvironment. We classified cell shapes into three categories and measured the extension of cells by fitting with an ellipsoid. From the trend of cell extensions, a cell in a large pore resembles one on 2D soft gel surface. This result suggests a crossover in length from 2D-like to 3D-like morphology for cell cultured on a curved surface.

並列關鍵字

3D culture ； 3T3 fibroblasts ； morphology ； autofluorescence ； gelatin scaffold ； monodisperse pores

參考文獻

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