CAJ | 학술논문

[目的] 分析经卵黄囊(YS)、胎肝(FL)和骨髓来源(BM)的基质细胞条件培养液(SCCM)诱导产生的胚胎干细胞(ES)来源造血细胞的细胞表型与功能差异. [方法] 制备YS-SCCM、 FL-SCCM及BM-SCCM,将3种基质细胞条件培养液分别加入ESE 14.1细胞分化培养体系培养7 d,通过对分化ESE14.1细胞造血发育表面标志FLK-1、Integrin-α4(整联蛋白-α4)、Sca-1(干细胞抗原-1)、CD34的检测、体外高增殖潜能集落形成细胞(HPP-CFC)分析及体内脾集落形成单位(CFU-S)检测,评价3种基质细胞条件培养液对ESE14.1细胞体外造血发育的调控作用.[结果] 经FL-SCCM诱导的EB细胞Flk-1+、Integrinα4+ 和Sca-1+ 细胞均高于YS-SCCM和BMSC-CM诱导组,分别为3.03%、2.9%和13.74%;经BMSC-CM诱导产生的CD34+ 细胞比例最高,为1.07% .经FLSC-CM或BMSC-CM诱导产生的造血细胞其HPP-CFC产率明显高于对照组,分别为7.4个/105细胞(P ＜ 0.01)和5.8个/105细胞(P ＜ 0.05);经FLSC-CM或BMSC-CM诱导产生的造血细胞其CFU-S产率亦明显高于对照组,分别为8.5个/5 × 105细胞和6.75个/5 × 105细胞(P ＜ 0.001). [结论] YS-SCCM、 FL-SCCM及BM-SCCM均可诱导ESE14.1向造血细胞分化,FL-SCCM和BM-SCCM造血定向诱导效率较高,所产生的细胞具备造血细胞的正常功能,FL-SCCM诱导产生的造血细胞原始程度高于BM-SCCM诱导产生的造血细胞.
[목적] 분석경란황낭(YS)、태간(FL)화골수래원(BM)적기질세포조건배양액(SCCM)유도산생적배태간세포(ES)래원조혈세포적세포표형여공능차이. [방법] 제비YS-SCCM、 FL-SCCM급BM-SCCM,장3충기질세포조건배양액분별가입ESE 14.1세포분화배양체계배양7 d,통과대분화ESE14.1세포조혈발육표면표지FLK-1、Integrin-α4(정련단백-α4)、Sca-1(간세포항원-1)、CD34적검측、체외고증식잠능집락형성세포(HPP-CFC)분석급체내비집락형성단위(CFU-S)검측,평개3충기질세포조건배양액대ESE14.1세포체외조혈발육적조공작용.[결과] 경FL-SCCM유도적EB세포Flk-1+、Integrinα4+ 화Sca-1+ 세포균고우YS-SCCM화BMSC-CM유도조,분별위3.03%、2.9%화13.74%;경BMSC-CM유도산생적CD34+ 세포비례최고,위1.07% .경FLSC-CM혹BMSC-CM유도산생적조혈세포기HPP-CFC산솔명현고우대조조,분별위7.4개/105세포(P ＜ 0.01)화5.8개/105세포(P ＜ 0.05);경FLSC-CM혹BMSC-CM유도산생적조혈세포기CFU-S산솔역명현고우대조조,분별위8.5개/5 × 105세포화6.75개/5 × 105세포(P ＜ 0.001). [결론] YS-SCCM、 FL-SCCM급BM-SCCM균가유도ESE14.1향조혈세포분화,FL-SCCM화BM-SCCM조혈정향유도효솔교고,소산생적세포구비조혈세포적정상공능,FL-SCCM유도산생적조혈세포원시정도고우BM-SCCM유도산생적조혈세포.
[Objective] To establish an effective and stable method to induce hematopoietic cells from embryonic stem(ES) cells,the phenotype and function of ES-derived hematopoietic cells induced by stromal cell conditioned medium (SCCM) of yolk sac (YS),fetal liver (FL) or bone marrow (BM) were analyzed and compared.[Methods] 10% of YS-SCCM,FL-SCCM or BM-SCCM was added to culture system for differentiation of ES cells.Flow cytometric analysis was used to identify expression of Flk1,Integrin α4,Sca-1,and CD34.Colony analysis was used to identify the quantity of high proliferative potential colony-forming cells (HPP-CFC) in differentiated ES cells.The yield of CFU-S (colony-forming unit-spleen) was also analyzed by transplanting ES cell derivatives into lethally irradiated mice.[Results] Expression of Flk1,Integrin α4,Sca-1,and CD34 could be tested on induced EB cells.The percentage of Flk-1+,Integrin α4+ and Sca-1+ cells induced by were 3.03%,2.9%,and 13.74%,respectively,which are greater than other groups.The percentage of CD34+ cells induced by BMSC-CM was 1.07% which was greater than other groups.The yields of HPP-CFC from hematopoietic cells induced by FLSC-CM or BMSC-CM were 7.4 /105 cells (P ＜ 0.01) and 5.8 /105 cells (P ＜ 0.05) which were greater than the yields of control group.The yields of CFU-S from hematopoietic cells induced by FLSC-CM or BMSC-CM were 8.5/5 × 105 cells and 6.75/5 × 105 cells which were also greater than the yields of control group (P ＜ 0.001).[Conclusion] Both YS-SCCM,FL-SCCM,and BM-SCCM could promote hematopoietic differentiation of ESE14.1 cells.Hematopoietic differentiation induced by FL-SCCM or BM-SCCM is more effective,which generates hematopoietic progenitor cells with normal function.Application of FL-SCCM generates more primitive hematopoietic progenitor cells than that of BM-SCCM.