探討豬羊水幹細胞於小鼠炎症性腸病模式之細胞治療與影響

The Cell-Based Therapeutic Effects of Stem Cells Derived from Amniotic Fluid of Pig on Experimental Colitis Mouse Models

林坤儀(Kun-Yi Lin)

指導教授 ： 吳信志

國立臺灣大學/生物資源暨農學院/生物科技研究所/博士(2017年)

https://doi.org/10.6342/NTU201704346

摘要

在現今運動醫學的領域裡，韌帶損傷一直是運動選手的夢魘。尤其在棒球場上，投手常常因為競賽重複投球而導致手肘的韌帶裂傷。我國近幾年旅外的著名投手，如郭泓志，陳偉殷等，都相繼因為手肘韌帶撕裂傷而分別接受手術或細胞注射治療。但是究竟是哪種治療為佳，關係著運動選手韌帶受傷的情形，以及再生醫學領域的進步程度。
身為一個手外科醫師，對於這種手肘尺外側副韌帶撕裂傷的病患，了解手術的預後，找尋新的治療方式來突破目前手術的限制是相當重要的。臨床統計結果顯示手術治療於第一級及第二級韌帶損傷之病患有較佳的預後，反觀第三級合併有內側副韌帶損傷的患者，其接受手術治療的效果較差。而且所有手術的病患皆出現術後肘關節僵硬，活動受限等情形。顯示臨床上手術治療仍造成目前無法解決的併發症。所以本臨床試驗先比較此韌帶受傷後造成的肘後外側旋轉不穩定之手術經驗，觀察病患的預後及併發症，進而探討幹細胞治療的可能性。
目前羊水來源前驅細胞已被證實具有多分化性 (pluripotency) 的潛力。所以在基礎試驗中我們利用本實驗室產製之攜帶紅色螢光蛋白轉基因豬與攜帶綠色螢光蛋白轉基因小鼠來建立羊水來源前驅細胞，由於之前實驗中，觀察到攜帶綠色螢光蛋白質轉基因小鼠之羊水來源前驅細胞，經由子宮內注射後，在腸道裡偵測到大部分的螢光訊號，所以本實驗使用攜帶紅色螢光蛋白質轉基因豬之羊水幹細胞，來治療腸道受損的小鼠。經由小鼠腹腔內注射羊水幹細胞一週後，可減少小鼠腸道受損長度，減輕腸道絨毛破壞程度，修復腸壁黏膜，並降低發炎反應，幫助小鼠腸道功能的恢復。因此羊水幹細胞具有遷移及修復腸道組織的能力。
了解如何建立羊水幹細胞，並觀察其治療小鼠受損腸道軟組織的潛力，期待未來能應用於臨床中韌帶及軟組織傷害之治療，使運動員更健康，拉長運動員的職業生涯，為運動醫學的治療領域帶來新的突破。

關鍵字

豬羊水幹細胞 ； 小鼠炎症性腸病模式

並列摘要

In the field of sports medicine, ligament injuries are usually players’ worst nightmare as they can often be career killers. Particularly in baseball, repetition of the throwing motion can cause ligament tears in the elbows for pitchers. Famous Taiwanese pitchers such as Hong-Chi Kuo and Wei-Yin Chen, both played in MLB, had respectively underwent surgeries and platelet-rich plasma injection due to their elbow ligament injuries. Still, the best treatment approach will depend on individual players’ specific ligament injuries and the development of regenerative medicine.
When treating patients with lateral ulnar collateral ligament tear, it is instrumental for a hand surgeon to understand the important of seeking post-op treatment to compliment the limitations of surgeries. Clinical statistics show that patients with stage I or stage II ligament injuries have better prognosis after surgeries while patients having stage III injuries with medial collateral ligament tears benefit less from surgical treatments. Moreover, all patients who underwent surgeries exhibit stiffness and limited mobility in the elbow joints. This indicates that the surgical approach has yet to resolve all complications at the moment. As such, the clinical study aims to compare the surgical results of ligament injuries caused by posterolateral rotatory instability of the elbow, observe patients’ post-op prognosis and complications, and then explore the possibility of stem-cell treatment.
As the multipotent potential of amniotic fluid-derived progenitor cells (AFPCs) has been proven, we used the red fluorescent protein (Ds-red)-expressing transgenic pigs and enhanced green fluorescent protein (EGFP)-labeled transgenic mice bred in our lab to produce AFPCs for tracing and tracking purpose in the pilot studies. Because most of the fluorescent signals were detected in the intestinal tract in intrauterine injections of AFPCs from EGFP transgenic mice in past studies, we use amniotic fluid stem cells (pAFSCs) from DsRed transgenic pigs to treat mice with intestinal injuries.
After one week of intraperitoneal injection of pAFSCs, we found that the injured intestines had been increased in length, some damages of the intestinal villi and intestinal walls had been repaired, and inflammation had been alleviated, which can all be contributory in improving the intestinal functions of the mice. Therefore, pAFSCs have the ability to move and repair intestinal tissues.
The study aims to find the best practice to produce pAFSCs and to observe their abilities to repair damaged soft intestinal tissues in mice, all in the hopes of applying the findings to the clinical treatment of ligament and soft tissue injuries, to improve players’ health and extend the life of their sports careers, and to bring about breakthroughs in the treatments of sports medicine.

並列關鍵字

porcine amniotic fluid-derived stem cells ； colitis mouse model

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