Hs68與HaCaT懸浮共培養的形態分析

Abstract

我們發展出一種共培養方法，主要是利用懸浮培養的方式進行共培養，有別於傳統作法，這種方式培養出的細胞會聚合成球，形成立體的構型，而比起其他類似的系統，譬如說懸滴培養法，或是水膠培養法，我們的方法更簡便，而且可以支持長時間的培養。 本研究採用的細胞是纖維母細胞與角質細胞的共培養，纖維母細胞使用的是Hs68細胞系，角質細胞使用的則是HaCaT細胞系。這兩株細胞系皆來自成年男性的包皮，並且也被證實此兩種細胞有交互作用發生。 由於Hs68和HaCaT有懸浮在幾丁聚醣涂佈的細胞培養盤上的特性，我們利用這點發展成一個懸浮共培養方法，在這個系統下，可以很間單的利用曠時攝影的方式觀察細胞的聚集。本研究討論了Hs68和HaCaT於懸浮共培養下細胞的聚集模式，並利用了簡單的模擬去分析。目前在實際操作中的已經有了初步的效果。 而學理上，我們採用了鈣黏蛋白的不同去解釋我們模型的設計，所以，我們也做了有關細胞凝聚力的分析，用來支持我們的模型。

We developed a new co-culture method mainly in co-culturing cells in suspension. Unlike the traditional method, cells cultured by this method will aggregate to a sphere and form a 3D structure, which compared to other systems like the hanging drop method and the hydrogel method, our system is simple, easy and good in long term culture. Our research used fibroblast and keratinocyte in co-culture, and we adopted Hs68 cell line from fibroblast and HaCaT cell line from keratinocyte. These cells are both from mature male foreskin, and the interaction existing between the two cells have been confirmed. We used the shared characteristics of Hs68 and HaCaT, that both will suspend in chitosan coated plate, to perform a suspension co-culture method. In this system, we can easily observe how each cells aggregate in time-lapse photography. This paper also discussed the aggregation process in suspension co-culture system, which was analyzed by a simple simulation. We’ve got a first step results in hands-on practice at present. Academically, we also adopted the theory about cadherin to explain our model design, thus we conducted some experiments in measuring cell-cell cohesion force to support our model.