本文選題：間充質(zhì)干細(xì)胞 + 免疫性肝損傷��； 參考：《河北醫(yī)科大學(xué)》2009年博士論文

【摘要】： 目的:近年來(lái),間充質(zhì)干細(xì)胞(mesenchymal stem cell,MSC)以其能在自體、同種異體甚至異種異體體內(nèi),旺盛地增殖和多向地分化,有效修復(fù)甚至完全重建功能?chē)?yán)重受損甚或幾近缺如的組織器官而不被排斥,已經(jīng)成為組織修復(fù)和再生重建領(lǐng)域內(nèi)備受關(guān)注的研究熱點(diǎn)。在人源MSC中,胎兒源MSC與成人源者相比,因其發(fā)育上更早、更少被病原體感染、安全性更高、獲取和制備相對(duì)更為捷易等優(yōu)點(diǎn),更是成為人們多愿選擇的來(lái)源。 現(xiàn)代免疫學(xué)研究已經(jīng)揭示,自體和同基因個(gè)體(如同卵孿生和純系內(nèi)個(gè)體)間的組織器官移植之所以不被排斥,是因供體與受體的組織相容性抗原一致或幾近一致所致。而同種異體間,尤其是異種異體間的組織器官移植一定會(huì)被排斥,是因供體和受體間組織相容性抗原不一致而致,且兩者間的組織相容性抗原相差越遠(yuǎn),則排斥反應(yīng)的發(fā)生就越迅速、越強(qiáng)烈。由是,在人類為修復(fù)或重建功能已嚴(yán)重受損甚至已幾近缺如的組織器官,除同卵孿生外,因?qū)嶋y覓組織相容性抗原一致或幾近一致的供體,不得已而在移植后必須長(zhǎng)期應(yīng)用昂貴的免疫抑制劑維持,但隨之又會(huì)帶來(lái)易發(fā)嚴(yán)重感染甚至腫瘤的危險(xiǎn)。由此,既不難理解為何人們會(huì)對(duì)人源尤其是胎兒源MSC寄予厚望而備受關(guān)注,同時(shí)也強(qiáng)烈提示,同種異體特別是異種異體MSC移植不被排斥,是否是其本身具有介導(dǎo)和啟動(dòng)有效免疫抑制作用機(jī)制所致;如是,則在其應(yīng)用之后是否有抑制機(jī)體免疫功能特別是招致腫瘤發(fā)生的風(fēng)險(xiǎn)性,勢(shì)必就成為該領(lǐng)域中必須予以高度重視和深入探索的重要課題。 迄今,已有許多研究小組對(duì)MSC在組織修復(fù)和免疫耐受重建等領(lǐng)域的實(shí)驗(yàn)研究和臨床應(yīng)用,進(jìn)行了多方面的大膽探索和勇敢嘗試。有關(guān)心肌修復(fù)、骨疾病及代謝性疾病的MSC細(xì)胞治療的臨床試驗(yàn)已在世界各地先后開(kāi)展;而Le Blanc及其同事應(yīng)用同種異體MSC于移植物抗宿主病(graft versus host disease, GVHD)的大膽嘗試,更堅(jiān)定了人們將其應(yīng)用于免疫損傷性疾病治療的信心。還有將自體或同種異體MSC以不同給藥方式(局部、靜脈和腹腔)治療實(shí)驗(yàn)性自身免疫性腦脊髓炎(experimental autoimmune encephalomyelitis, EAE)及Ⅱ型膠原誘導(dǎo)的類風(fēng)濕性關(guān)節(jié)炎(collagenⅡ-induced arthritis, CIA)模型的研究均獲成功。更兼我們驚奇地發(fā)現(xiàn)經(jīng)與未經(jīng)預(yù)處理的人胎兒源MSC,對(duì)ConA誘導(dǎo)的免疫性肝損傷有雙效作用,因這確是在世界上尚無(wú)報(bào)道,而極大地激發(fā)了我們對(duì)人胎兒源MSC免疫生物學(xué)特性及其與腫瘤免疫學(xué)關(guān)系的濃厚研究興趣。 為此,本課題分離鑒定了胎兒骨髓源和羊水源MSC以優(yōu)化來(lái)源,研究其免疫抑制功能特別是對(duì)ConA誘導(dǎo)免疫性肝損傷的雙效作用,在分析其介導(dǎo)的免疫抑制分子的基礎(chǔ)上,探索其之致瘤的風(fēng)險(xiǎn)性,為MSC的進(jìn)一步研究和應(yīng)用提供新的理論和實(shí)驗(yàn)依據(jù)。 方法:貼壁消化傳代培養(yǎng)法,自人胎兒骨髓及羊水細(xì)胞獲取,并經(jīng)形態(tài)學(xué)、細(xì)胞表面標(biāo)記和分化潛能鑒定,確系為人胎兒源MSC(以下簡(jiǎn)寫(xiě)為hMSC)。MTS法評(píng)價(jià)hMSC對(duì)ConA誘導(dǎo)小鼠脾細(xì)胞增殖的抑制作用。獲取ConA刺激小鼠脾細(xì)胞增殖的培養(yǎng)上清,以其培養(yǎng)hMSC而獲取經(jīng)預(yù)處理的hMSC;觀察其生長(zhǎng)增殖、細(xì)胞表型、染色體核型等細(xì)胞生物學(xué)特性。參照Tiegs等的方法建立ConA誘導(dǎo)免疫性肝損傷動(dòng)物模型,肝損傷指標(biāo)包括小鼠存活率、血清谷丙轉(zhuǎn)氨酶(glutamate-pyruvate transaminase,ALT)和谷草轉(zhuǎn)氨酶(glutamic-oxalacetic transaminase,AST)含量以及肝組織病理?yè)p傷情況等。以模型動(dòng)物肝損傷指標(biāo)為對(duì)照,觀察經(jīng)或未經(jīng)預(yù)處理的hMSC對(duì)模型動(dòng)物肝損傷的影響,評(píng)價(jià)其對(duì)肝損傷的保護(hù)或促進(jìn)作用。裸鼠皮下移植,排除hMSC成瘤可能;選擇人肝癌細(xì)胞系MHCC/97H腫瘤細(xì)胞和小鼠肝癌細(xì)胞系H22腫瘤細(xì)胞為對(duì)象,觀察hMSC對(duì)腫瘤細(xì)胞體內(nèi)/外細(xì)胞周期、體外克隆形成能力、荷瘤小鼠生存期、皮下移植瘤體內(nèi)生長(zhǎng)增殖以及體內(nèi)/外侵襲轉(zhuǎn)移等的影響,評(píng)價(jià)其致瘤的風(fēng)險(xiǎn)性。 結(jié)果: 1人胎兒骨髓源及羊水源MSC的分離鑒定及其免疫抑制作用 貼壁消化傳代培養(yǎng)法,自人胎兒骨髓及羊水細(xì)胞獲取,連續(xù)傳代培養(yǎng)3~5代后,其形態(tài)學(xué)、細(xì)胞表面標(biāo)記和分化潛能鑒定,均具人MSC典型特征,確系獲得了合格的hMSC。 ConA刺激小鼠脾細(xì)胞增殖實(shí)驗(yàn)顯示,hMSC有顯著的、劑量依賴式的免疫抑制作用(P0.05)。 2未預(yù)處理的hMSC對(duì)ConA誘導(dǎo)小鼠免疫性肝損傷的作用 未預(yù)處理的hMSC可顯著降低ConA誘導(dǎo)免疫性肝損傷模型小鼠存活率(P0.05)、升高模型小鼠血清中肝功能指標(biāo)ALT和AST含量(P0.05)并促進(jìn)肝組織的病理?yè)p傷(P0.05)。顯示未預(yù)處理的hMSC對(duì)ConA誘導(dǎo)免疫性肝損傷有顯著的促進(jìn)作用。 3未預(yù)處理的hMSC對(duì)模型小鼠肝損傷的促進(jìn)作用與HLA分子表達(dá)及補(bǔ)體C3介導(dǎo)的關(guān)系 HLA-A,B,C(+)/HLA-DR(-)及HLA-A,B,C(-)/HLA-DR(-)的hMSC均可顯著升高模型小鼠血清中肝損傷指標(biāo)ALT的含量(P0.05),顯示未預(yù)處理的hMSC對(duì)模型小鼠肝損傷的促進(jìn)作用,與其表面HLA分子表達(dá)無(wú)關(guān);各組小鼠血清ALT和補(bǔ)體C3含量測(cè)定及回歸分析,顯示未預(yù)處理的hMSC促進(jìn)模型小鼠肝損傷亦與補(bǔ)體C3介導(dǎo)無(wú)關(guān)。 4未預(yù)處理的hMSC對(duì)正常小鼠肝臟組織的影響 小鼠肝組織病理切片HE染色結(jié)果顯示,未預(yù)處理的hMSC組與正常組小鼠相比,肝小葉周邊、界板肝細(xì)胞及匯管區(qū)的點(diǎn)灶狀溶解性壞死、凋亡小體及匯管區(qū)炎等病理指標(biāo)差異明顯(P0.05);提示未預(yù)處理的hMSC腹腔注射可致正常小鼠肝組織輕微病理?yè)p傷。 5經(jīng)預(yù)處理的hMSC對(duì)ConA誘導(dǎo)小鼠免疫性肝損傷的作用。 經(jīng)預(yù)處理的hMSC可顯著升高ConA誘導(dǎo)免疫性肝損傷模型小鼠存活率(P0.05)、降低模型小鼠血清中肝功能指標(biāo)ALT含量(P0.05)并顯著降低肝組織的病理?yè)p傷程度(P0.05);顯示經(jīng)預(yù)處理的hMSC對(duì)ConA誘導(dǎo)免疫性肝損傷有顯著的保護(hù)作用。 6經(jīng)預(yù)處理的hMSC對(duì)模型小鼠肝組織FasL表達(dá)和脾Treg亞群細(xì)胞比例的影響。 經(jīng)預(yù)處理的hMSC可顯著減少模型組小鼠肝組織FasL表達(dá)(P0.05)并增加早期時(shí)相(3h)脾細(xì)胞Treg亞群細(xì)胞比例(P0.05)。提示使模型鼠肝組織FasL表達(dá)減少和脾Treg細(xì)胞比例增加,可能是經(jīng)預(yù)處理的hMSC對(duì)模型小鼠免疫性肝損傷具有保護(hù)作用的分子機(jī)制。 7經(jīng)與未經(jīng)預(yù)處理的hMSC相關(guān)細(xì)胞因子分泌差異 經(jīng)預(yù)處理的hMSC其IL-6、IL-8及RANTES等細(xì)胞因子分泌,較未預(yù)處理的hMSC增多。提示,經(jīng)預(yù)處理的hMSC有可能藉增加這些細(xì)胞因子的分泌,參與對(duì)ConA誘導(dǎo)免疫性肝損傷的保護(hù)作用。 8經(jīng)預(yù)處理的hMSC的自身成瘤性 以ConA刺激小鼠脾細(xì)胞增殖的培養(yǎng)上清對(duì)hMSC行預(yù)處理后, hMSC的細(xì)胞周期被阻于G0/G1期,增殖指數(shù)顯著降低(P0.05);細(xì)胞表型和染色體核型保持不變;接種于裸鼠皮下,亦無(wú)腫瘤形成。提示預(yù)處理未導(dǎo)致hMSC的自身成瘤性。 9經(jīng)預(yù)處理的hMSC的促瘤性 經(jīng)與未經(jīng)預(yù)處理的hMSC,均可使人肝癌細(xì)胞系MHCC/97H腫瘤細(xì)胞及小鼠肝癌細(xì)胞系H22腫瘤細(xì)胞阻滯于G0/G1期,顯著降低兩種腫瘤細(xì)胞的增殖指數(shù)(P0.05);均可使MHCC/97H腫瘤細(xì)胞體外克隆形成能力顯著降低(P0.05);預(yù)處理與否,不影響hMSC對(duì)這兩種腫瘤細(xì)胞發(fā)揮上述這些抑制作用的強(qiáng)度(P0.05)。經(jīng)與未經(jīng)預(yù)處理的hMSC,對(duì)MHCC/97H腫瘤細(xì)胞和H22腫瘤細(xì)胞體內(nèi)移植瘤的生長(zhǎng)增殖、體內(nèi)/外侵襲轉(zhuǎn)移及荷瘤小鼠生存期無(wú)影響(P0.05)。提示預(yù)處理未導(dǎo)致hMSC的促瘤性。 結(jié)論: 1貼壁消化傳代培養(yǎng)法可有效從骨髓和羊水,獲取合格的人胎兒源MSC。 2人胎兒源MSC對(duì)ConA刺激的T細(xì)胞增殖有顯著的免疫抑制作用。 3經(jīng)或未經(jīng)ConA刺激T細(xì)胞增殖的培養(yǎng)上清預(yù)處理,可顯著改變?nèi)颂涸碝SC對(duì)ConA誘導(dǎo)免疫性肝損傷的影響模式,此結(jié)果與結(jié)論國(guó)內(nèi)外未見(jiàn)報(bào)道。 4未預(yù)處理的人胎兒源MSC對(duì)ConA誘導(dǎo)免疫性肝損傷有顯著的促進(jìn)作用。 5經(jīng)預(yù)處理的人胎兒源MSC對(duì)ConA誘導(dǎo)免疫性肝損傷有顯著的保護(hù)作用。 6未預(yù)處理的人胎兒源MSC對(duì)ConA誘導(dǎo)免疫肝損傷的促進(jìn)作用與其表面HLA分子表達(dá)及補(bǔ)體C3的介導(dǎo)作用無(wú)關(guān)。 7經(jīng)預(yù)處理的人胎兒源MSC對(duì)ConA誘導(dǎo)免疫肝損傷的保護(hù)作用與其致肝組織FasL表達(dá)降低及脾細(xì)胞早期時(shí)相(3h)Treg亞群細(xì)胞比例增高有關(guān)。 8經(jīng)預(yù)處理的人胎兒源MSC,其IL-6、IL-8及RANTES細(xì)胞因子表達(dá)增加。 9人胎兒源MSC在觀察期內(nèi)未見(jiàn)成瘤性和促瘤活性,初步判斷其在所觀察期內(nèi)不具有致瘤風(fēng)險(xiǎn)性。
[Abstract]:Objective: in recent years, mesenchymal stem cell (MSC) has become a field of tissue repair and regenerative reconstruction with its ability to proliferate and multidirectional differentiation in autologous, allogeneic and heterogeneous allogeneic bodies, to effectively repair and even completely reconstruct the tissues and organs that have been severely damaged or almost absent. In human source MSC, fetal source MSC, compared with adult source, has the advantages of earlier development, less infection by pathogens, higher safety, and relatively easy access to preparation and preparation, and is also the source of people's choice.
Modern immunological studies have revealed that tissue and organ transplantation between autologous and homologous individuals (like egg twins and individuals in pure lines) is not excluded because of the consistent or close agreement of the donor and receptor histocompatibility antigen, and the allograft, especially the allogeneic allograft, will be rejected. Due to the inconsistency of the histocompatibility antigen between the donor and the recipient, and the farther the difference of the histocompatibility antigen between the two is, the more rapid and stronger the rejection occurs. A consistent or nearly consistent donor must not be used for long-term application of expensive immunosuppressive agents, but it will also bring about the risk of severe infection or even cancer. Thus, it is not difficult to understand why people are highly concerned about the expectations of human sources, especially fetal MSC, and also strongly suggest that the allograft is special. Not heterologous allograft MSC transplantation is not rejected, whether it has the mechanism to mediate and activate the effective immunosuppressive mechanism. If, if it has the risk of inhibiting the body's immune function, especially the occurrence of tumor, it is bound to be an important subject in this field.
So far, many research teams have made bold and bold attempts at the experimental research and clinical application of MSC in the fields of tissue repair and immune tolerance reconstruction. The clinical trials of MSC cell therapy for myocardial repair, bone disease and metabolic diseases have been carried out all over the world; and Le Blanc and its colleagues should A bold attempt to use allogeneic MSC for graft versus host disease (GVHD), and to strengthen its confidence in the treatment of immune injury diseases, and the treatment of experimental autoimmune encephalomyelitis (experimental auto) by the use of autologous or allogenic MSC in different ways (local, vein and abdominal cavity) Immune encephalomyelitis, EAE) and type II collagen induced rheumatoid arthritis (collagen II -induced arthritis, CIA) have been successfully studied. Moreover, we were surprised to find that the untreated human fetal MSC, which has double effects on the immune liver injury induced by ConA, is not yet reported in the world. The earth has stimulated our interest in studying the immunological characteristics of human fetal MSC and its relationship with tumor immunology.
Therefore, we isolated and identified the fetal bone marrow source and sheep water source MSC to optimize the source and study the double effect of its immunosuppressive function, especially on ConA induced immune liver injury. On the basis of the analysis of its mediated immunosuppressive molecules, the risk of its tumor induced tumor was explored, and a new theory for further research and application of MSC was provided. Experimental basis.
Methods: human fetal bone marrow and amniotic fluid cells were obtained from human fetal bone marrow and amniotic fluid cells, and were identified by morphology, cell surface labeling and differentiation potential. The inhibitory effect of hMSC on the proliferation of spleen cells induced by ConA was evaluated by human fetal MSC (below hMSC).MTS method. The culture supernatant of spleen cell proliferation stimulated by ConA was obtained. HMSC was cultured and pretreated hMSC was obtained, and the cell biological characteristics of its growth, proliferation, cell phenotype, chromosome karyotype and so on were observed. The animal models of immune liver injury induced by ConA were established with reference to Tiegs, including the survival rate of the mice, the serum alanine transaminase (ALT), and the transglutaminate of cereal grass. Enzyme (glutamic-oxalacetic transaminase, AST) content as well as pathological damage of liver tissue. The effect of hMSC on liver injury of model animals was observed or untreated by the model animal liver damage index, and the protective or promoting effect on liver injury was evaluated. The subcutaneous transplantation of nude mice, the possibility of removing hMSC into the tumor, was used to select human liver cancer. The cell line MHCC/97H tumor cells and the mouse liver cancer cell line H22 tumor cells were used to observe the effects of hMSC on the cell / outer cell cycle of the tumor cells, the ability of in vitro cloning and formation, the survival period of the tumor bearing mice, the growth and proliferation of the subcutaneous transplanted tumor and the invasion and metastasis of the body / outside, and to evaluate the risk of the tumor.
Result:
Isolation, identification and immunosuppressive effect of MSC from 1 human fetal bone marrow and sheep water sources
After being obtained from human fetal bone marrow and amniotic fluid cells from human fetal bone marrow and amniotic fluid cells, after continuous subculture of 3~5 generation, the morphology, cell surface labeling and differentiation potential were identified. All of them have the typical characteristics of human MSC, and the qualified hMSC. is obtained.
ConA stimulated splenocytes proliferation in mice showed that hMSC had a significant dose-dependent immunosuppressive effect (P0.05).
2 the effect of untreated hMSC on ConA induced immune liver injury in mice
Untreated hMSC could significantly reduce the survival rate of ConA induced immune liver injury model mice (P0.05), increase the liver function index ALT and AST content (P0.05) in the serum of the model mice and promote the pathological damage of liver tissue (P0.05). It showed that the untreated hMSC had a significant effect on the immune liver injury induced by ConA.
3 the effect of untreated hMSC on liver injury in mice and the expression of HLA and complement C3.
HLA-A, B, C (+) /HLA-DR (-) and HLA-A, B, C (-) /HLA-DR (-) hMSC can significantly increase the content of the liver damage index ALT content (P0.05) in the serum of the model mice, showing that the unpretreated hMSC is not related to the expression of the liver damage in the model mice. The treatment of hMSC promoted liver damage in model mice was also independent of complement C3 mediating.
4 Effect of untreated hMSC on liver tissue in normal mice
The results of HE staining in the pathological section of liver tissue in mice showed that compared with the normal group, the hMSC group of the untreated group was significantly different from the normal group of the liver lobules, the focal dissolving necrosis of the focal liver cells and the sink area, the apoptotic bodies and the manifold inflammation (P0.05), suggesting that the untreated hMSC could lead to a slight liver tissue in normal mice. Pathological injury.
5 the effect of pretreated hMSC on ConA induced immune liver injury in mice.
The pretreated hMSC significantly increased the survival rate of ConA induced immune liver injury model mice (P0.05), reduced the liver function index ALT content (P0.05) in the serum of the model mice and significantly reduced the degree of pathological damage of liver tissue (P0.05), and showed that the pretreated hMSC had a significant protective effect on ConA induced immune liver injury.
6 the effect of pretreated hMSC on the expression of FasL and the proportion of Treg subsets in liver tissue of model mice.
The pretreated hMSC could significantly reduce the FasL expression of liver tissue in the model mice (P0.05) and increase the Treg subgroup ratio (P0.05) in the early phase (3H) of the spleen cells (P0.05), suggesting that the reduced expression of FasL in the liver tissue of the model mice and the proportion of the spleen Treg cells increased, which may be a protective molecule of the pretreated hMSC on the immune liver damage in the model mice. Mechanism.
7 differences in hMSC related cytokine secretion between pretreated and untreated
The pretreated hMSC, such as IL-6, IL-8 and RANTES, is more secreted than untreated hMSC. It is suggested that the pretreated hMSC may be involved in the protection of ConA induced immune liver injury by increasing the secretion of these cytokines.
8 pretreated hMSC's self tumorigenicity
The cell cycle of hMSC was pretreated with ConA stimulated the proliferation of mouse splenocytes, the cell cycle of hMSC was blocked at G0/G1, the proliferation index decreased significantly (P0.05), the cell phenotype and chromosome karyotype remained unchanged, and no tumor was formed in subcutaneous inoculation of nude mice, suggesting that preconditioning did not lead to the self tumorigenicity of hMSC.
9 pretreated hMSC's tumorigenicity
The hMSC tumor cells of human hepatoma cell line MHCC/97H and H22 tumor cell line H22 were blocked at G0/G1 stage, and the proliferation index of two tumor cells was significantly reduced (P0.05), and the ability of MHCC/97H tumor cells to be cloned in vitro was significantly reduced (P0.05). The preconditioning did not affect hMSC against this two. The growth and proliferation of tumor cells in MHCC/97H tumor cells and H22 tumor cells, in vivo / out invasion and metastasis and the survival period of tumor bearing mice (P0.05) were not affected by pretreated hMSC (P0.05). It suggested that the preconditioning did not lead to the tumor growth of hMSC in the tumor cells (P0.05).
Conclusion:
1 adherent digestion and subculture can effectively obtain MSC. from human bone marrow and amniotic fluid.
The 2 fetal fetal MSC has significant immunosuppressive effect on the proliferation of T cells stimulated by ConA.
3 the preconditioning of cultured supernatant with or without ConA stimulation of T cell proliferation can significantly change the model of human fetal source MSC on ConA induced immune liver injury. The results and conclusions have not been reported at home and abroad.
4 untreated human fetal MSC has a significant effect on ConA induced immune liver injury.
5 pretreated human fetal MSC has a significant protective effect on ConA induced immune liver injury.
6 the effect of unpretreated human fetal MSC on ConA induced liver injury is not related to the expression of HLA on the surface and the mediating effect of complement C3.
7 the protective effect of pretreated human fetal MSC on ConA induced immune liver injury is related to the decrease of FasL expression in liver tissue and the increase in the proportion of Treg subgroup cells in the early phase of the spleen cells (3H).
8 the expression of IL-6, IL-8 and RANTES cytokines increased in pretreated human fetal MSC.
There was no tumorigenic and tumorigenic activity in the 9 fetal fetal MSC during the observation period.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R575;R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 ;Cytokines,STATs and Liver Disease[J];Cellular & Molecular Immunology;2005年02期

2 ;Transplantation of Human Bone Marrow Mesenchymal Stem Cell Ameliorates the Autoimmune Pathogenesis in MRL/lpr Mice[J];Cellular & Molecular Immunology;2008年06期

3 ;Immunosuppressive effects of rat mesenchymal stem cells:involvement of CD4~+ CD25~+ regulatory T cells[J];Hepatobiliary & Pancreatic Diseases International;2008年06期

4 胡潔;王秀榮;齊義新;劉運(yùn)江;;乳腺癌白細(xì)胞介素-8基因表達(dá)的臨床檢測(cè)意義[J];河北醫(yī)科大學(xué)學(xué)報(bào);2006年05期

5 李萬(wàn)勝;胡潔;趙娟;魏林;程建新;;IL-8基因真核表達(dá)載體的構(gòu)建及其在OVCAR-3卵巢癌細(xì)胞中的表達(dá)[J];軍事醫(yī)學(xué)科學(xué)院院刊;2008年02期

6 胡潔;齊義新;姚智燕;魏林;;乳腺癌患者白細(xì)胞介素8臨床檢測(cè)的意義[J];臨床薈萃;2006年10期

7 Patrizia Bossolasco;Tiziana Montemurro;Lidia Cova;Stefano Zangrossi;Cinzia Calzarossa;Simona Buiatiotis;Davide Soligo;Silvano Bosari;Vincenzo Silani;Giorgio Lambertenghi Deliliers;PaoloRebulla;LorenzaLazzari;;Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential[J];Cell Research;2006年04期

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