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
2008年
51期
10129-10133
,共5页
方建培%翁文骏%吴燕峰%杨兴鸽%许吕宏%徐宏贵%魏菁
方建培%翁文駿%吳燕峰%楊興鴿%許呂宏%徐宏貴%魏菁
방건배%옹문준%오연봉%양흥합%허려굉%서굉귀%위정
脐血%造血干细胞%地中海贫血%群体反应性抗体%半固体集落培养
臍血%造血榦細胞%地中海貧血%群體反應性抗體%半固體集落培養
제혈%조혈간세포%지중해빈혈%군체반응성항체%반고체집락배양
背景:已有报道证实群体反应性抗体在实体器官移植受者排斥反应中起着重要作用,并且与植入物功能衰竭呈正相关.目的:实验拟观察反复输血地中海贫血患儿血清中特异性群体反应性抗体对脐血造血干/祖细胞增殖、分化能力的影响.设计、时间及地点:体外细胞学实验,于2006-01/2007-08在中山大学附属第二医院医学研究中心完成.材料:5份健康足月产妇脐带血,每份80-100 mL.反复输血地中海贫血患儿的群体反应性抗体血清样本.5份健康儿童AB分型血清样本.6份阳性抗凝静脉血样本10 mL.方法:采用聚蔗糖-泛影葡胺梯度离心法分离脐带血单个核细胞.将1×105脐血单个核细胞与分别与健康儿童AB血清50 μ L、0,50,100μL不同浓度群体反应性抗体血清混合后,再将上述细胞混合物与兔补体共同孵育进行半固体集落培养.主要观察指标:倒置显微镜下观察并计数共培养第7,14天总集落数和各种集落数.结果:脐血造血干/祖细胞受群体反应性抗体血清作用后,其半固体集落培养的总集落数以及各种集落数均有不同程度的下降,其中总集落数和粒单系集落形成单位的下降数,经统计学处理差异具有显著性意义(P<0.01):群体反应性抗体血清量与总集落、粒单系集落形成单位、粒红单核巨核系集落形成单位、红系爆式集落形成单位、巨核系爆式集落形成单位、巨核系集落形成单位之间的肯德尔相关分析呈负相关,经统计学处理差异有显著性意义(P<0.05).结论:特异性群体反应性抗体对脐血造血干/祖细胞的增殖、分化能力具有抑制作用,并ⅱ且抑制作用与群体反应性抗体剂量相关:群体反应性抗体剂量越大,抑制作用越强.
揹景:已有報道證實群體反應性抗體在實體器官移植受者排斥反應中起著重要作用,併且與植入物功能衰竭呈正相關.目的:實驗擬觀察反複輸血地中海貧血患兒血清中特異性群體反應性抗體對臍血造血榦/祖細胞增殖、分化能力的影響.設計、時間及地點:體外細胞學實驗,于2006-01/2007-08在中山大學附屬第二醫院醫學研究中心完成.材料:5份健康足月產婦臍帶血,每份80-100 mL.反複輸血地中海貧血患兒的群體反應性抗體血清樣本.5份健康兒童AB分型血清樣本.6份暘性抗凝靜脈血樣本10 mL.方法:採用聚蔗糖-汎影葡胺梯度離心法分離臍帶血單箇覈細胞.將1×105臍血單箇覈細胞與分彆與健康兒童AB血清50 μ L、0,50,100μL不同濃度群體反應性抗體血清混閤後,再將上述細胞混閤物與兔補體共同孵育進行半固體集落培養.主要觀察指標:倒置顯微鏡下觀察併計數共培養第7,14天總集落數和各種集落數.結果:臍血造血榦/祖細胞受群體反應性抗體血清作用後,其半固體集落培養的總集落數以及各種集落數均有不同程度的下降,其中總集落數和粒單繫集落形成單位的下降數,經統計學處理差異具有顯著性意義(P<0.01):群體反應性抗體血清量與總集落、粒單繫集落形成單位、粒紅單覈巨覈繫集落形成單位、紅繫爆式集落形成單位、巨覈繫爆式集落形成單位、巨覈繫集落形成單位之間的肯德爾相關分析呈負相關,經統計學處理差異有顯著性意義(P<0.05).結論:特異性群體反應性抗體對臍血造血榦/祖細胞的增殖、分化能力具有抑製作用,併ⅱ且抑製作用與群體反應性抗體劑量相關:群體反應性抗體劑量越大,抑製作用越彊.
배경:이유보도증실군체반응성항체재실체기관이식수자배척반응중기착중요작용,병차여식입물공능쇠갈정정상관.목적:실험의관찰반복수혈지중해빈혈환인혈청중특이성군체반응성항체대제혈조혈간/조세포증식、분화능력적영향.설계、시간급지점:체외세포학실험,우2006-01/2007-08재중산대학부속제이의원의학연구중심완성.재료:5빈건강족월산부제대혈,매빈80-100 mL.반복수혈지중해빈혈환인적군체반응성항체혈청양본.5빈건강인동AB분형혈청양본.6빈양성항응정맥혈양본10 mL.방법:채용취자당-범영포알제도리심법분리제대혈단개핵세포.장1×105제혈단개핵세포여분별여건강인동AB혈청50 μ L、0,50,100μL불동농도군체반응성항체혈청혼합후,재장상술세포혼합물여토보체공동부육진행반고체집락배양.주요관찰지표:도치현미경하관찰병계수공배양제7,14천총집락수화각충집락수.결과:제혈조혈간/조세포수군체반응성항체혈청작용후,기반고체집락배양적총집락수이급각충집락수균유불동정도적하강,기중총집락수화립단계집락형성단위적하강수,경통계학처리차이구유현저성의의(P<0.01):군체반응성항체혈청량여총집락、립단계집락형성단위、립홍단핵거핵계집락형성단위、홍계폭식집락형성단위、거핵계폭식집락형성단위、거핵계집락형성단위지간적긍덕이상관분석정부상관,경통계학처리차이유현저성의의(P<0.05).결론:특이성군체반응성항체대제혈조혈간/조세포적증식、분화능력구유억제작용,병ⅱ차억제작용여군체반응성항체제량상관:군체반응성항체제량월대,억제작용월강.
BACKGROUND: Panel reaction antibody (PRA) plays an important role in rejection of recipients undergoing solid organ transplantation, which has a positive effect on nonfunction of implant. OBJECTIVE: To evaluate the effect of thalassemic serum-specific PRA on the proliferation and differentiation of umbilical cord blood hernatopoetic stem/progenitor cells (HSC/HPCs) in children patients with thalassemia. DESIGN, TIME AND SETTING: The in vitro cytology experiment was performed at the Experimental Research Center, Second Affiliated Hospital, Zhongshan University from January 2006 to August 2007. MATERIALS: Five samples of umbilical cord blood from healthy full-term birth puerperants (each 80 100 mL) were used in this study. PRA serum samples of children patients with thalassemia after repetitive blood transfusion, five samples of AB blood grouping serum, and six samples of positive anticoagulation vein blood (10 mL) were used in the study. METHODS: Mononuclear cells were harvested from umbilical cord blood by Ficoll-Hypaque gradient centrifugation. 1 × 105 rnononuclear cells from umbilical cord blood were incubated with different levels of experimental or AB control serum (0, 50, 100 μ L) from healthy children. The mixture mentioned above was incubated with rabbit complement for semisolid colony culture.MAIN OUTCOME MEASURES: Colony-forming units (CFU) were counted and observed after 7 days and 14 days of culture under an inverted microscope.RESULTS: After incubation with HSC/HPCs PRA serum, total number of CFUs and varied CFUs decreased to different extents, of which the total number of CFUs and CFU- granulocyte-rnacrophages (CFU-GM) had significant differences (P < 0.01). Moreover, there were negative correlations between different levels of serum PRA and the followings: number of total colonies, CFU- GM, CFU- granulocyte-erythrocyte-monocyte-megakaryocytes, CFU-erythroids, burst forming unit-megakaryocytes, and CFU-megakaryocytes (P < 0.05).CONCLUSION: The thalassemic serum PRA has an apparent inhibitory effect on the proliferation and differentiation of cord blood HSC/HPCs in vitro, an effect that may be pronounced with increasing serum PRA.