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The myocardial differentiation ability from Adipose - Derived Adult Stem cells on chitosan films
rat adipose - derived adult stem cells(ADASs)
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|摘要:||本研究是先利用不同生醫材料－TCPS(T)、PVA(P)、Chitosan 1(CS1)、Chitosan 2(CS 2)來探討大鼠脂肪幹細胞 (rADASs) 經過3天成球 + 1天5-aza有無誘導 + 7天分化(共11天的分化時間)，來比較心肌分化的能力，由心肌基因表現可看出Chitosan 2的分化能力為最加佳。更進一步，藉由CBD-RGD結合 Chitosan 2 所改質的材料- CS2R，進行11天的分化培養後，發現CBD-RGD在第一天的貼附功效，使細胞聚集成的細胞球比CS2更大，而由心肌基因表現和Tight junction (ZO-1)免疫螢光染色發現CBD-RGD確實提升了CS2的分化能力，且有加5-aza誘導的心肌分化能力又為最好；也由心肌免疫螢光染色實驗發現有添加5-aza的CS2與CS2R的表現幾乎都比無添加的組別好；再由倒立顯微鏡與SEM圖可發現， 5-aza的添加會減少細胞從細胞球跑出的比例。因此，大細胞球的聚集與5-aza確實增強了心肌的分化能力。|
In this study, the myocardial differentiation ability of rat Adipose-Derived Adult Stem cells (rADAS) had first been compared by culturing rADAS on different biomaterials including Chitosan 1 (CS 1), Chitosan 2 (CS 2), PVA (P), and TCPS (T). The rADAS culturing condition proceeded with 3 days for spheroids formation, 1 day for 5-aza induction, and 7 days for differentiation (a total of 11 days of differentiation time). The results of the RT-PCR assay showed rADAS cultured on CS2 had better myocardial differentiation ability than rADAS cultured on other biomaterials. Further, by modifying CS2 with CBD-RGD (CS2R), it had been observed that the attachment effect of CBD-RGD could induce bigger spheroid formations than the spheroids formed on CS2. From the RT-PCRassay and immunocytochemical staining of tight junction, CBD-RGD indeed could improve myocardial differentiation ability of rADAS. And as shown from conventional light microscope and SEM pictures, CS2R with 5-aza induction could decrease rADAS migration out of spheroids,and consequently, ensure higher myocardial differentiation. Therefore, CS2R with 5-aza induction together could induce the myocardial differentiation of rADAS better than CS2R without 5-aza induction.
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