北京生物醫(yī)學(xué)工程

灌注法大鼠全肺臟脫細(xì)胞基質(zhì)支架的建立

Whole lung extracellular matrix scaffold in rats by perfusion

作者：黑飛龍周建業(yè) 羅富良吳蓓陳蒙蒙楊勝男龍村

單位：                      中國醫(yī)學(xué)科學(xué)院  北京協(xié)和醫(yī)學(xué)院阜外心血管病醫(yī)院體外循環(huán)科（北京100037）        

關(guān)鍵詞：組織工程；肺臟；脫細(xì)胞基質(zhì)

分類號(hào)：

出版年·卷·期（頁碼）：2013·32·1（17-21）

摘要：

目的應(yīng)用灌注法建立大鼠全肺臟脫細(xì)胞基質(zhì)支架，探討灌注法制備的肺脫細(xì)胞基質(zhì)作為細(xì)胞支架構(gòu)建組織工程器官的可行性。方法 12周齡Wistar大鼠20只，麻醉后完整切取肺組織。由肺動(dòng)脈插入18G灌注頭連接到Langendorff灌注平臺(tái)，保持灌注壓40cmH2O。肺臟用肝素化PBS液將殘留血液沖洗干凈，然后用1%脫氧膽酸鈉溶液灌注90min，再用核酸酶應(yīng)用液灌注30min。檢測指標(biāo)包括常規(guī)蘇木素伊紅(HE)染色觀察細(xì)胞形態(tài)；Weigert彈力纖維+Von Gieson結(jié)締組織(ET+VG)染色觀察膠原纖維和彈性纖維的分布。結(jié)果進(jìn)行脫細(xì)胞灌注后，大鼠肺臟逐漸呈均一的白色半透明狀，顯微鏡觀察肺組織細(xì)胞已基本消失，而膠原纖維排列較整齊，彈性纖維保存良好。結(jié)論灌注法是一種簡單有效構(gòu)建大鼠全肺組織基質(zhì)支架的技術(shù)，1%脫氧膽酸鈉溶液能夠較好地脫除大鼠肺臟的細(xì)胞。

Objectives To produce whole-lung extracellular matrix(ECM) scaffold in rats by perfusion and to assess the feasibility of ECM as the cytoskeleton and tissue-engineered organ construction. Methods Lungs were harvested from twenty 12-week-old Wistar rats.Lungs were explanted，cannulated with an 18G cannula，and perfused (constant pressure，40cmH2O) through the pulmonary artery on Langendorff system with heparinized phosphate-buffered saline (PBS)，1% sodium deoxycholate in deionized water (90min) and RNase & DNase away (30min).HE staining was performed to confirm the removal of cells and Von Gieson staining wasperformed to show the integrity of collagen and elastin.Results After the decellularization process,rat lung scaffolds showed white color and looked semitransparent.The cells were removed effectively from lung tissue,while the collagen and elastin were kept intact structure in the scaffolds.Conclusions Perfusion technic is simple and effective to produce rat whole-lung ECM, and 1% sodium deoxycholate solution can better remove rat lung cells.

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