中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2006年
29期
160-163
,共4页
赖颖晖%赖永榕%卢玉英%莫武宁
賴穎暉%賴永榕%盧玉英%莫武寧
뢰영휘%뢰영용%로옥영%막무저
造血干细胞%流式细胞术%细胞移植%小鼠
造血榦細胞%流式細胞術%細胞移植%小鼠
조혈간세포%류식세포술%세포이식%소서
背景:造血干细胞来源目前包括骨髓干细胞、外周血干细胞和脐带血干细胞,寻找新的干细胞来源以满足临床移植需要一直是人们的希望.从妊娠第5周起,肝脏中发育成完善的血窦系统,此后造血干细胞就可以随血液流动迁移.目的:观察人胎儿血造血干/祖细胞的生物学特性并对其进行非肥胖糖尿病/重症联合免疫缺陷小鼠移植.设计:对照实验.单位:广西医科大学第一附属医院血液科.对象:①细胞来源:21例胎儿血标本取自胎龄为18～29周[(24.2±3.2)周]死亡胎儿及21例足月脐带血标本取自广西医科大学第一附属医院产科2002-10/2003-02.所取的血标本均取得其家属的知情同意.②实验动物:非肥胖糖尿病/重症联合免疫缺陷小鼠12只,6～7周龄,雌性,无菌饲养于超净工作台中.方法:采用流式细胞术,检测胎儿血的造血干/祖细胞表面标志,包括CD34,CD38,HLA-DR及CD90,同时与21例足月儿脐血作比较.并将人胎血单个核细胞移植给6只经亚致死量照射的非肥胖糖尿病/重症联合免疫缺陷小鼠,5周后观察其植入情况,采用流式细胞术检测小鼠骨髓中人白细胞的含量,以及采用聚合酶链反应检测小鼠骨髓中的人Cart-1基因.主要观察指标:①胎血和脐血造血干/祖细胞表面标志的表达情况.②将胎血细胞移植给非肥胖糖尿病/重症联合免疫缺陷小鼠的植入情况.结果:①人胎血中CD34+细胞的百分率显著高于足月脐血[(2.258 8±0.720 9)%,(1.572 9±0.478 3)%,P=0.000 4],CD34+CD38-细胞和CD34+CD90+细胞的百分率也均显著高于足月脐血[(1.298 6±0.470 6)%,(0.871 0±0.409 5)%,P=0.0016;(0.9300±0.469 2)%,(0.5600±0.365 8)%.P=0.032 4].②6例胎血中的4例可顺利重建经亚致死量照射的非肥胖糖尿病/重症联合免疫缺陷小鼠的造血,移植后5周在小鼠骨髓中仍可检测到人的白细胞和人Cart-1基因.结论:人胎儿血中有比足月脐血更高含量的造血干/祖细胞,其单个核细胞能植入非肥胖糖尿病/重症联合免疫缺陷小鼠骨髓,并重建髓、淋巴系全面造血.胎儿血有望成为多能造血干细胞来源.
揹景:造血榦細胞來源目前包括骨髓榦細胞、外週血榦細胞和臍帶血榦細胞,尋找新的榦細胞來源以滿足臨床移植需要一直是人們的希望.從妊娠第5週起,肝髒中髮育成完善的血竇繫統,此後造血榦細胞就可以隨血液流動遷移.目的:觀察人胎兒血造血榦/祖細胞的生物學特性併對其進行非肥胖糖尿病/重癥聯閤免疫缺陷小鼠移植.設計:對照實驗.單位:廣西醫科大學第一附屬醫院血液科.對象:①細胞來源:21例胎兒血標本取自胎齡為18～29週[(24.2±3.2)週]死亡胎兒及21例足月臍帶血標本取自廣西醫科大學第一附屬醫院產科2002-10/2003-02.所取的血標本均取得其傢屬的知情同意.②實驗動物:非肥胖糖尿病/重癥聯閤免疫缺陷小鼠12隻,6～7週齡,雌性,無菌飼養于超淨工作檯中.方法:採用流式細胞術,檢測胎兒血的造血榦/祖細胞錶麵標誌,包括CD34,CD38,HLA-DR及CD90,同時與21例足月兒臍血作比較.併將人胎血單箇覈細胞移植給6隻經亞緻死量照射的非肥胖糖尿病/重癥聯閤免疫缺陷小鼠,5週後觀察其植入情況,採用流式細胞術檢測小鼠骨髓中人白細胞的含量,以及採用聚閤酶鏈反應檢測小鼠骨髓中的人Cart-1基因.主要觀察指標:①胎血和臍血造血榦/祖細胞錶麵標誌的錶達情況.②將胎血細胞移植給非肥胖糖尿病/重癥聯閤免疫缺陷小鼠的植入情況.結果:①人胎血中CD34+細胞的百分率顯著高于足月臍血[(2.258 8±0.720 9)%,(1.572 9±0.478 3)%,P=0.000 4],CD34+CD38-細胞和CD34+CD90+細胞的百分率也均顯著高于足月臍血[(1.298 6±0.470 6)%,(0.871 0±0.409 5)%,P=0.0016;(0.9300±0.469 2)%,(0.5600±0.365 8)%.P=0.032 4].②6例胎血中的4例可順利重建經亞緻死量照射的非肥胖糖尿病/重癥聯閤免疫缺陷小鼠的造血,移植後5週在小鼠骨髓中仍可檢測到人的白細胞和人Cart-1基因.結論:人胎兒血中有比足月臍血更高含量的造血榦/祖細胞,其單箇覈細胞能植入非肥胖糖尿病/重癥聯閤免疫缺陷小鼠骨髓,併重建髓、淋巴繫全麵造血.胎兒血有望成為多能造血榦細胞來源.
배경:조혈간세포래원목전포괄골수간세포、외주혈간세포화제대혈간세포,심조신적간세포래원이만족림상이식수요일직시인문적희망.종임신제5주기,간장중발육성완선적혈두계통,차후조혈간세포취가이수혈액류동천이.목적:관찰인태인혈조혈간/조세포적생물학특성병대기진행비비반당뇨병/중증연합면역결함소서이식.설계:대조실험.단위:엄서의과대학제일부속의원혈액과.대상:①세포래원:21례태인혈표본취자태령위18～29주[(24.2±3.2)주]사망태인급21례족월제대혈표본취자엄서의과대학제일부속의원산과2002-10/2003-02.소취적혈표본균취득기가속적지정동의.②실험동물:비비반당뇨병/중증연합면역결함소서12지,6～7주령,자성,무균사양우초정공작태중.방법:채용류식세포술,검측태인혈적조혈간/조세포표면표지,포괄CD34,CD38,HLA-DR급CD90,동시여21례족월인제혈작비교.병장인태혈단개핵세포이식급6지경아치사량조사적비비반당뇨병/중증연합면역결함소서,5주후관찰기식입정황,채용류식세포술검측소서골수중인백세포적함량,이급채용취합매련반응검측소서골수중적인Cart-1기인.주요관찰지표:①태혈화제혈조혈간/조세포표면표지적표체정황.②장태혈세포이식급비비반당뇨병/중증연합면역결함소서적식입정황.결과:①인태혈중CD34+세포적백분솔현저고우족월제혈[(2.258 8±0.720 9)%,(1.572 9±0.478 3)%,P=0.000 4],CD34+CD38-세포화CD34+CD90+세포적백분솔야균현저고우족월제혈[(1.298 6±0.470 6)%,(0.871 0±0.409 5)%,P=0.0016;(0.9300±0.469 2)%,(0.5600±0.365 8)%.P=0.032 4].②6례태혈중적4례가순리중건경아치사량조사적비비반당뇨병/중증연합면역결함소서적조혈,이식후5주재소서골수중잉가검측도인적백세포화인Cart-1기인.결론:인태인혈중유비족월제혈경고함량적조혈간/조세포,기단개핵세포능식입비비반당뇨병/중증연합면역결함소서골수,병중건수、림파계전면조혈.태인혈유망성위다능조혈간세포래원.
BACKGROUND: Currently the hematopoietic stem cells can be obtained from bone marrow, peripheral blood and cord blood, so it is expected to search a new source of stem cells in order to satisfy the clinical transplantation needs. From the 5th week of pregnancy, the blood sinusoid system develops completely in liver, and then hematopoietic stem cells can move with blood flow. OBJECTIVE: To observe the biological features of human fetal blood hematopoietic stem/progenitor cells (HS/PCs), and their transplantation into non-obese diabetic/severe combined immunodeficiency disease (NOD/ SCID) mice. DESIGN: Control trial. SETTING: Department of Hematology, First Affiliated Hospital of Guangxi Medical University. MATERIALS:①Cell resource: Twenty-one fetal blood samples were from dead fetus [gestational age of 18-29 weeks, mean (24.2±3.2) weeks] and twenty-one full-term cord blood samples were provided from the Department of Obstetrics, First Affiliated Hospital of Guangxi Medical University between October 2002 and February 2003, with the consent of their relatives.②Experimental animal: Twelve NOD/SCID female mice of 6-7 weeks old were bred in sterility and super-clean operation board. METHODS: Flow cytometer was used to assess cell surface markers of HS/PCs including CD34, CD38, HLA-DR and CD90 in 21 human fetal blood samples, and their expressions were compared with 21 human cord blood samples. Moreover, human fetal blood mononuclear cells (MNCs) were transplanted into 6 NOD/SCID mice irradiated sublethally. After 5 weeks, human leukocytic content was also detected in bone marrow of mice with flow cytometer while human Cart-1 gene in recipients' bone marrow was sensed with polymerase chain reaction (PCR).MAIN OUTCOME MEASURES: ① Expressions of HS/PCs surface markers in fetal blood and cord blood. ②Implantation of fetal blood cells into NOD/SCID mice.RESULTS: ①The percentage of CD34+ cells in fetal blood was significantly higher than that of full-term cord blood [(2.258 8±0.720 9)%,(1.572 9±0.478 3)%, P=0.000 4]. The percentages of CD34+CD38- cells and CD34+CD90+ cells in fetal blood were also higher than those of fullterm cord blood [(1.298 6±0.470 6)%, (0.871 0±0.409 5)%, P=0.001 6;(0.930 0±0.469 2)%, (0.560 0±0.365 8)%, P=0.032 4].②Four cases (4/6)of human fetal blood MNCs smoothly transplanted the hematopoiesis of sublethally irradiated NOD/SCID mice. Five weeks after the transplantation, human leukocyte and Cart-1 gene could still be detected in marrow cells of NOD/SCID mice.CONCLUSION: Human fetal blood contains more HS/PCs than cord blood. Human fetal blood MNCs can engraft bone marrow of NOD/SCID mice and reconstitute general hemopoiesis of marrow and lymph systems.Human fetal blood is a new possible source of pluripotential hematopoietic stem cell.
