中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
37期
7397-7400
,共4页
岑妍慧%林江%何国珍%王进声%陈青%林涌%赵小芳%黄荣师%单华%冯祖德%叶家良%邓绍策
岑妍慧%林江%何國珍%王進聲%陳青%林湧%趙小芳%黃榮師%單華%馮祖德%葉傢良%鄧紹策
잠연혜%림강%하국진%왕진성%진청%림용%조소방%황영사%단화%풍조덕%협가량%산소책
马氏珠母贝%细胞培养%角质细胞生长因子
馬氏珠母貝%細胞培養%角質細胞生長因子
마씨주모패%세포배양%각질세포생장인자
背景:关于消化外套膜组织获得的细胞悬液是否能在体外有效地扩增形成珍珠囊以及最终形成珍珠仍存在争议,且这方面研究不多.目的:建立一套有效的马氏珠母贝外套膜组织细胞体外分离及培养的技术和方法,同时探讨体外形成具有完整结构和分泌功能的珍珠囊并最终生成优质珍珠的最佳方法.设计、时间及地点:单一样本观察,于2008-08/12在广西中医学院基础医学院完成.材料:贝龄1.0～2.0岁马氏珠母贝由广西北海市营盘珍珠实业有限公司提供,自配改进的海水贝类平衡盐溶液(MWBSS);自制珍珠贝血清和珍珠贝体液;角质细胞生长因子为美国Sigma公司产品.方法:使用2.5 g/L胰蛋白酶消化马氏珠母贝外套膜组织,收获的细胞使用M199(含体积分数10%胎牛血清)培养基进行培养,并在其中加入10 μg/L的角质细胞生长因子、10%自制的珍珠贝体液和贝血清,持续培养30 d.主要观察指标:细胞生长特性和生长状态.结果:体外培养的珍珠外套膜上皮细胞增殖迅速,分泌功能旺盛,肌肉细胞增殖能力强,并最终能包裹外套膜上皮细胞,形成结构较完整、分泌能力较强的珍珠囊.结论:使用改进的培养技术和培养体系中添加角质细胞生长因子在体外培养珍珠外套膜组织细胞,可获得生长状态和分泌功能均较好的珍珠囊.
揹景:關于消化外套膜組織穫得的細胞懸液是否能在體外有效地擴增形成珍珠囊以及最終形成珍珠仍存在爭議,且這方麵研究不多.目的:建立一套有效的馬氏珠母貝外套膜組織細胞體外分離及培養的技術和方法,同時探討體外形成具有完整結構和分泌功能的珍珠囊併最終生成優質珍珠的最佳方法.設計、時間及地點:單一樣本觀察,于2008-08/12在廣西中醫學院基礎醫學院完成.材料:貝齡1.0～2.0歲馬氏珠母貝由廣西北海市營盤珍珠實業有限公司提供,自配改進的海水貝類平衡鹽溶液(MWBSS);自製珍珠貝血清和珍珠貝體液;角質細胞生長因子為美國Sigma公司產品.方法:使用2.5 g/L胰蛋白酶消化馬氏珠母貝外套膜組織,收穫的細胞使用M199(含體積分數10%胎牛血清)培養基進行培養,併在其中加入10 μg/L的角質細胞生長因子、10%自製的珍珠貝體液和貝血清,持續培養30 d.主要觀察指標:細胞生長特性和生長狀態.結果:體外培養的珍珠外套膜上皮細胞增殖迅速,分泌功能旺盛,肌肉細胞增殖能力彊,併最終能包裹外套膜上皮細胞,形成結構較完整、分泌能力較彊的珍珠囊.結論:使用改進的培養技術和培養體繫中添加角質細胞生長因子在體外培養珍珠外套膜組織細胞,可穫得生長狀態和分泌功能均較好的珍珠囊.
배경:관우소화외투막조직획득적세포현액시부능재체외유효지확증형성진주낭이급최종형성진주잉존재쟁의,차저방면연구불다.목적:건립일투유효적마씨주모패외투막조직세포체외분리급배양적기술화방법,동시탐토체외형성구유완정결구화분비공능적진주낭병최종생성우질진주적최가방법.설계、시간급지점:단일양본관찰,우2008-08/12재엄서중의학원기출의학원완성.재료:패령1.0～2.0세마씨주모패유엄서북해시영반진주실업유한공사제공,자배개진적해수패류평형염용액(MWBSS);자제진주패혈청화진주패체액;각질세포생장인자위미국Sigma공사산품.방법:사용2.5 g/L이단백매소화마씨주모패외투막조직,수획적세포사용M199(함체적분수10%태우혈청)배양기진행배양,병재기중가입10 μg/L적각질세포생장인자、10%자제적진주패체액화패혈청,지속배양30 d.주요관찰지표:세포생장특성화생장상태.결과:체외배양적진주외투막상피세포증식신속,분비공능왕성,기육세포증식능력강,병최종능포과외투막상피세포,형성결구교완정、분비능력교강적진주낭.결론:사용개진적배양기술화배양체계중첨가각질세포생장인자재체외배양진주외투막조직세포,가획득생장상태화분비공능균교호적진주낭.
BACKGROUND: There is a great debate but little research addressing the cell suspension obtained from the digested mantle tissues can effective amplify and form pearl sac in vitro, thus producing pearl. OBJECTIVE: To establish an effective technique and method of in vitro separation and culture of mantle of the pearl oyster (Pinctada martensii), and to determine the optimal method of forming pearl sac with the intact structure and secretion function, thus producing pearl. DESIGN, TIME AND SETTING: Single sample observation was performed at the School of Basic Medicine, Guangxi Traditional Chinese Medical University, between August and December in 2008. MATERIALS: Pearl oyster (Pinctada martensii) aged 1.0 2.0 years, were offered by Yingpan Pearl Industrial Co., Ltd. Of Beihai City, China; the self-modified marine shellfish balanced salt solution; the self-prepared concha pteriae serum and concha pteriae body fluid; keratinocyte growth factor was purchased from Sigma, USA. METHODS: The mantle of pearl oyster (Pinctada martensii) was digested with 2.5 g/L trypsin, the harvested cells were cultured using M199 medium containing 10% fetal bovine serum and supplemented with 10 μg/L keratinocyte growth factor, 10% self-prepared concha pteriae serum and concha pteriae body fluid. The cultivation was performed for 30 days. MAIN OUTCOME MEASURES: Cell growth characteristics and growth state. RESULTS: The pearl mantle epithelial cells cultured in vitro were shown to proliferate rapidly, secrete productively, and the muscle cells showed a great proliferation, finally encapsulated the mantle epithelial cells to form pear sac with the intact structure and strong secretion function. CONCLUSION: Using the modified culture technology and culture system, the addition of keratinocyte growth factor can obtain the well growing and secreting pearl sac during in vitro culture of mantle cells.
