干細(xì)胞的基本知識(shí)

1、干細(xì)胞(stem cell),,干細(xì)胞(stem cell),干細(xì)胞是具有自我更新與分化潛能的未分化或低分化細(xì)胞。醫(yī)學(xué)界稱(chēng)之為“萬(wàn)用細(xì)胞”,干細(xì)胞具有“無(wú)限” 的增殖能力，能夠產(chǎn)生與母代細(xì)胞相同的子代細(xì)胞，維持該干細(xì)胞種群。,自我更新,分化潛能,干細(xì)胞能分化生成不同表型的成熟細(xì)胞，例如胚胎干細(xì)胞可以分化形成個(gè)體的所有成熟細(xì)胞類(lèi)型。,根據(jù)其發(fā)育階段不同，可分為：,胚胎干細(xì)胞 (ESC)具有分化為機(jī)體任何一種組織器官的潛能。

2、 如囊胚期內(nèi)細(xì)胞團(tuán)中的細(xì)胞。成體干細(xì)胞（ASC)機(jī)體組織中保留的一部分比較原始的細(xì)胞,一旦機(jī)體需要,這些細(xì)胞便可按發(fā)育途徑,先分裂后分化,產(chǎn)生分化細(xì)胞。,按分化潛能的大小,全能干細(xì)胞： 在一定條件下，能夠分化發(fā)育成為完整個(gè)體的細(xì)胞，如哺乳動(dòng)物桑葚胚的8細(xì)胞期之前的細(xì)胞 。多能干細(xì)胞： 在胚胎發(fā)育的三胚層形成后，細(xì)胞的分化潛能受到限制，僅能向本胚層組織和器官方向分化發(fā)育。如造血多能干細(xì)胞。

3、單能干細(xì)胞:只能分化為一種類(lèi)型的細(xì)胞，而且自我更新能力有限，如上皮組織基底層的干細(xì)胞，肌肉中的成肌細(xì)胞，紅系干細(xì)胞等。,骨髓干細(xì)胞,一類(lèi)是造血干細(xì)胞（hematopoietic stem cells，HSCs），它為循環(huán)血液提供前體細(xì)胞；另一類(lèi)是間充質(zhì)干細(xì)胞（mesenchymal stem cells，MSCs），它是骨髓中主要的 支持細(xì)胞，在調(diào)節(jié)造血干細(xì)胞的長(zhǎng)期存活，生長(zhǎng)分化中起重要作用。,二、干細(xì)胞的生物學(xué)特征,（一） 具有

4、“無(wú)限”的自我更新能力,（二）多向分化潛能,（三）具有未分化或低分化特性 (四) 可塑性,,,,1、干細(xì)胞的獨(dú)特增殖方式,干細(xì)胞通過(guò)非對(duì)稱(chēng)分裂產(chǎn)生與一個(gè)與母代細(xì)胞完全相同的子代細(xì)胞，以保持干細(xì)胞穩(wěn)定；同時(shí)還產(chǎn)生分化細(xì)胞。,2.細(xì)胞分化的潛能隨個(gè)體發(fā)育進(jìn) 程逐漸“縮窄”,細(xì)胞分化的一般規(guī)律：在胚胎發(fā)育過(guò)程中，細(xì)胞逐漸由“全能”到“多能”，最后向“單能”的趨向。,干細(xì)胞的可塑性,干細(xì)胞的可塑性：即干細(xì)胞具有在不同微環(huán)境中可轉(zhuǎn)化

5、為不同干細(xì)胞及不同類(lèi)型的成熟組織細(xì)胞的特性。機(jī)制：1, 橫向分化 2，去分化/脫分化 3，干細(xì)胞的異質(zhì)性 4，細(xì)胞融合 5，基因重組學(xué)說(shuō) 6，亞全能干細(xì)胞學(xué)說(shuō)影響因素：微環(huán)境和基因表達(dá),,,三 鑒定，篩選,ESC:胚胎細(xì)胞在發(fā)育的不同階段, 其細(xì)胞表面出現(xiàn)不同的抗原。未分化的人ESC 細(xì)胞表面S

6、SEA-3、SSEA-4、TI-1-60、TRA-1-81等呈陽(yáng)性.這些標(biāo)記物加上干細(xì)胞時(shí)期細(xì)胞內(nèi)堿性磷酸酶、端粒酶的特異性高表達(dá), 可成為鑒定胚胎干細(xì)胞的依 據(jù).ASC的鑒定識(shí)別主要通過(guò)三種途徑: 分離培養(yǎng)和形態(tài)學(xué)觀(guān)察; 免疫表型鑒定; 分化功能檢測(cè)。,四 干細(xì)胞的應(yīng)用,一，干細(xì)胞是早期胚胎發(fā)育的良好模型二，干細(xì)胞是研究人類(lèi)疾病的良好模型三，干細(xì)胞具有臨床應(yīng)用的前景。1，移植治療2，基因治療􀀁3

7、，轉(zhuǎn)運(yùn)載體4，干細(xì)胞庫(kù),五 存在的問(wèn)題,1)干細(xì)胞的分離、純化、增殖、鑒定的問(wèn)題。 2)干細(xì)胞誘導(dǎo)分化的問(wèn)題。 3）移植時(shí)機(jī)的把握 4）供體細(xì)胞的功能表達(dá)問(wèn)題 5)移植后免疫排斥反應(yīng)。 6) 干細(xì)胞治療中生物安全，倫理和法律問(wèn)題,骨髓干細(xì)胞在抗肝纖維化治療中的應(yīng)用研究,優(yōu)點(diǎn)：1，近年來(lái)有不少文獻(xiàn)報(bào)道，骨髓干細(xì)胞可以分化為肝細(xì)胞，改善肝功能。2，研究發(fā)現(xiàn)，將肝星狀細(xì)胞與MSC 共培養(yǎng)后，受活性肝星狀細(xì)胞分泌的IL- 6

8、 刺激，MSC 分泌IL- 10、TNF-α 和肝細(xì)胞生長(zhǎng)因子（hepatocyte growth factor，HGF），使得肝星狀細(xì)胞增殖激活明顯受抑制，凋亡增加，膠原合成顯著減少。 3，相關(guān)資料顯示，MSCs 除了有可以分化為肝細(xì)胞的潛能外，還可以通過(guò)表達(dá)高水平的MMPs，尤其是MMP- 9來(lái)減少肝臟細(xì)胞外基質(zhì)的沉積。,弊端：1，最近研究顯示，BMSCs 移植入肝硬化小鼠體內(nèi)，骨髓源性肝細(xì)胞僅占肝組織細(xì)胞總數(shù)的0.6%，而骨

9、髓源性肝星狀細(xì)胞和肌成纖維細(xì)胞則分別高達(dá)68%和70%，且是通過(guò)非融合方式直接來(lái)源于BMSCs，并具有Ⅰ型膠原轉(zhuǎn)錄活性，能促進(jìn)纖維變性反應(yīng)。2，另外，也有學(xué)者認(rèn)為，慢性肝損傷還可誘導(dǎo)BMSCs 分化為肌成纖維細(xì)胞的前體細(xì)胞———成纖維細(xì)胞，潛在地促進(jìn)肝纖維化發(fā)展。因此用于肝再生的BMSCs 移植研究必須警惕其加重肝纖維化形成的可能。3，最值得引起注意的是骨髓干細(xì)胞可以促進(jìn)肝細(xì)胞性肝癌的發(fā)生。,The Therapeutic Pote

10、ntial of Human UmbilicalMesenchymal Stem Cells from Wharton’s Jellyin the Treatment of Rat Liver Fibrosis,Pei-Chun Tsai,1 Tz-Win Fu,5 Yi-Ming Arthur Chen,2 Tsui-Ling Ko,8 Tien-Hua Chen,3 Yang-Hsin Shih,6,9 Shih-Chieh H

11、ung,4,7* and Yu-Show Fu3,1,Abstract,We investigated the effect of human umbilical mesenchymal stem cells (HUMSCs) from Wharton’s jelly on carbon tetrachloride(CCl4)–induced liver fibrosis in rats. Rats were treated with

12、 CCl4 for 4 weeks, and this was followed by a direct injection of HUMSCs into their livers. After 4 more weeks of CCl4 treatment (8 weeks in all), rats with HUMSC transplants [CCl4(8W) ＋HUMSC liver] exhibited a significa

13、nt reduction in liver fibrosis, as evidenced by Sirius red staining and a collagen content assay, in comparison with rats treated with CCl4 for 8 weeks without HUMSC transplants [CCl4 (8W)]. Moreover, rats in theCCl4 (8

14、W)HUMSC (liver) group had significantly lower levels of serum glutamic oxaloacetic transaminase, glutamic pyruvate transaminase, a-smooth muscle actin, and transforming growth factor-1 in the liver, whereas the expressio

15、n of hepatic mesenchymal epithelial transition factor–phosphorylated type (Met-P) and hepatocyte growth factor was up-regulated, in comparison with the CCl4 (8W) group。,Notably, engrafted HUMSCs scattered mostly in the h

16、epatic connective tissue but did not differentiate into hepatocytes expressing human albumin or a -fetoprotein. Instead, these engrafted, undifferentiated HUMSCs secreted a variety of bioactive cytokines that may restor

17、e liver function and promote regeneration. Human cytokine assay revealed that the amounts of human cutaneous T cell–attracting chemokine, leukemia inhibitory factor, and prolactin were substantially greater in the livers

18、 of the CCl4 (8W)HUMSC (liver) group, with considerably reduced hepatic inflammation manifested by a micro positron emission tomography scan. Our findings suggest that xenogeneic transplantation of HUMSCs is a novel app

19、roach for treating liver fibrosis and may be a promising therapeutic intervention in the future.,Aim,We investigated the effect of human umbilical mesenchymal stem cells (HUMSCs) from Wharton’s jelly on carbon tetrachlor

20、ide(CCl4)–induced liver fibrosis in rats.,Materials and Methods,Rat:1,normal 2, CCl4 for 8 weeks 3, CCl4(8W) ＋HUMSC index: weight, CTACK, Prolactin, LIF ,Met-P HGF/GAPGH

21、 GOT,GPT, a-SMA, TGF-β1, collagen [18F]-FDG human albumin , human a-FP , fresh liver,Results,Conclusions,1,HUMSC transplantation can increase CTACK, Prolactin, LIF ,Met-P ,HGF/GAPGH

22、 and reduce GOT,GPT, a-SMA, TGF-β1, inflammation so that it can inhibit HSC proliferation and collagen synthesis and enhance liver cell repair.2, The effect of HUMSCs on reducing fibrogenesis most likely relies on bioac

23、tive factors or cytokines released from the grafted HUMSCs to trigger liver regeneration rather than on the differentiation of these cells into hepatocytes.In conclusion, sufficient amounts of HUMSCs in rat livers can s