靜水壓力和細胞外基質硬度對於肌肉母細胞分化之影響

Biophysical Cues on Modulation of Myoblast Differentiation Using Hydrostatic Pressure and Elastomeric Matrix of Tunable Stiffness

林鴻儒(Hung-Ju Lin)

指導教授 ： 郭柏齡

共同指導教授 ： 陳明豐(Ming-Fong Chen)

國立臺灣大學/電機資訊學院/生醫電子與資訊學研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.01055

摘要

細胞在體內存活、更新增生、和分化，是需要一個複雜且動態調控生長因子、細胞外基質、和生物物理力學刺激整合的微環境。因此本實驗目的欲探討結合壓力與細胞外基質硬度的生物物理刺激之下，對於肌肉母細胞分化的影響。
我們使用PA水膠 (polyacrylamide hydrogel)製程方式，利用acrylamide和bis-acrylamide的組成比例不同，來調整PA水膠硬度。再用Sulfo-SANPAH將fibronectin連接於PA水膠表面，以利於H9C2肌肉母細胞附著而生長分化。相對於控制組培養在35mm培養皿中，實驗組則是將H9C2肌肉母細胞於外加靜水壓力10 cmH2O培養48小時。然後用細胞螢光染色方式測定肌肉細胞分化轉錄因子，MyoD和myogenin，表現量在控制組和實驗組之間的差異。根據實驗結果，我們發現H9C2肌肉母細胞附著在不同細胞外基質硬度下，施予外加靜水壓力10 cmH2O皆有促進表現分化轉錄因子，MyoD和myogenin，的傾向。因此實驗結果可能暗示肌肉母細胞在培養過程中，附著於肌肉組織基質硬度範圍的肌肉母細胞，給予適當壓力刺激， 可能會促進肌肉細胞分化和成熟過程。
累積越來越多單一生物化學及物理性訊息刺激對於細胞分化成長影響的知識後，也讓以組織工程的方式來整合細胞所需動態或靜態的分化生長訊息來調控細胞分化及成熟過程和效率的可行性逐漸增加。因此建立一套兩種以上細胞分化生長訊息交互作用的研究方式，對於未來再生醫學和體外細胞實驗可有所助益。

關鍵字

靜水壓力 ； 水膠 ； 基質硬度 ； 肌肉母細胞 ； 生物物理

並列摘要

Accumulating evidence has demonstrated that integrated biochemical and biophysical microenvironmental cues with accurate temporal and spatial arrangements are needed in cell growth, self-renewal, and differentiation. Although muscle tissue pressure exists during the process of adult injury-induced myogenesis, a dearth of studies showed whether muscle regeneration would be altered when compressive pressure was applied during myoblast differentiation. Thus, the purpose of this study was to investigate the effects of hydrostatic pressure on differentiation of myoblasts, which was cultured on extracellular matrix of different stiffness to simulate the condition of embryonic myogenesis.
We cultured H9C2 myoblasts on fibronectin-coated polyacrylamide hydrogel, of which the stiffness was adjusted by different combinations of acrylamide and bis-acrylamide. Hydrostatic pressure of 10 cmH2O was applied. Then, we examined the expression of myogenesis transcriptional factors, MyoD and myogenin, using immunofluorescence staining. The results revealed that hydrostatic pressure increased the expression of MyoD and myogenin during myoblast differentiation. It might imply that hydrostatic pressure could facilitate the process of myogenesis. Therefore, further studies are merited on the detailed signaling pathways of pressure-induced modulation in myogenesis for developing muscle tissue engineering.

並列關鍵字

hydrostatic pressure ； myoblast differentiation ； myogenesis ； extracellular matrix ； stiffness ； polyacrylamide hydrogel ； biophysical cues

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