
浏览全部资源
扫码关注微信
重庆医科大学附属第一医院心内科
纸质出版日期:2011,
网络出版日期:2011-11-30,
移动端阅览
熊挺淋, 张晓刚, 赵霞, 等. 诱导多能干细胞/原代心肌细胞的融合细胞表现出双向重建[J]. 陆军军医大学学报, 2011,33(22):2376-2380.
XIONG TINGLIN, ZHANG XIAOGANG, ZHAO XIA, et al. Bidirectional reprogramming of fusion cells of pluripotent stem cells/primary cardiac myocytes. [J]. Journal of army medical university, 2011, 33(22): 2376-2380.
熊挺淋, 张晓刚, 赵霞, 等. 诱导多能干细胞/原代心肌细胞的融合细胞表现出双向重建[J]. 陆军军医大学学报, 2011,33(22):2376-2380. DOI:
XIONG TINGLIN, ZHANG XIAOGANG, ZHAO XIA, et al. Bidirectional reprogramming of fusion cells of pluripotent stem cells/primary cardiac myocytes. [J]. Journal of army medical university, 2011, 33(22): 2376-2380. DOI:
目的 体外构建诱导多能干细胞(induced pluripotent stem cells
iPSc)与原代心肌细胞的融合细胞,初步探讨融合细胞体外生物学特性。 方法 绿色荧光蛋白(green fluorescent protein
GFP)转染八聚体结合转录因子4(octamer-binding transcription factor-4, Oct-4)小鼠来源的iPSc与小鼠乳鼠心肌细胞通过聚乙二醇(polyethylene glycol, PEG-4000)诱导融合,建立细胞融合模型。动态观察融合细胞生长形态变化情况,融合细胞碱性磷酸酶(alkali phosphatase
AKP)染色,免疫荧光鉴定融合细胞表达干细胞与心肌细胞特异性蛋白情况,以及染色体核型分析,确定融合细胞是否发生细胞核的融合及程度。 结果 聚乙二醇(PEG-4000)能够介导iPSc与心肌细胞融合,融合后第4天开始出现成集落状生长的细胞团;第2、3、4、5 天的iPSc和融合细胞AKP阳性率分别为(0.935±0.039)、(0.939±0.022)、(0.954±0.017)、(0.944±0.027)和(0.761±0.044)、(0.740±0.023)、(0.681±0.034)、(0.748±0.045),同时间点的iPSc和融合细胞AKP阳性率比较有明显差异(P<0.05);融合细胞初期主要表现为iPSc的特点,Oct-4表达阳性,cTnT表达阴性,融合7 d后,逐渐表现出两种细胞的特点,Oct-4与cTnT均表达阳性;超过80%的杂交细胞染色体数目在76~80条之间。 结论 二倍体iPSc与二倍体心肌细胞的融合细胞,具有两亲本细胞的特性,表现出双向重建。
Objective To construct fusion cells with induced pluripotent stem cells (iPSc) and primary cardiac myocytes
in vitro
and to investigate biological features of the fusion cells. Methods Polyethylene glycol (PEG-4000) was used to mediate the cell fusion of iPSc
derived from green fluorescent protein (GFP) transgenes (octamer-binding transcription factor-4
Oct-4) mouse and cardiac myocytes from neonatal mouse. Morphological changes of the fusion cells were observed dynamically after alkaline phosphatase (AKP) staining. Specific proteins of stem cells and cardiac myocytes in fusion cells were detected by immunofluorescence. Chromosome karyotype analysis were performed to determine whether the occurrence of nuclear fusion and degree of integration. Results Fusion cells were constructed successfully by polyethylene glycol mediation. Colony-like cell clusters appeared in 4 d after fusion. The AKP positive rate of iPSc were 0.935±0.039
0.939±0.022
0.954±0.017
and 0.944±0.027 at the 2nd
3rd
4th and 5th days respectively
and that of fusion cells were 0.761±0.044
0.740±0.023
0.681±0.034
and 0.748±0.045 at the corresponding days respectively. At the same time points
there were significant differences between iPSc AKP-positive rates and those of fusion cells (
P
<
0.05). In the initial stage
fusion cells mainly displayed iPSc characteristics
with Oct-4 positive while cTnT negative. Then the fusion cells began to display both characteristics of iPSc and cardiac myocytes in 7 d after fusion
with positive expression of Oct-4 and cTnT. More than 80% of fusion cells had 76 to 80 chromosomes. Conclusion Fusion cells from diploid iPSc and diploid myocardial cells display the characteristics of the two parental cells and show bidirectional reprogramming.
细胞融合诱导多能干细胞心肌细胞融合细胞心肌梗死
cell fusioninduced pluripotent stem cellscardiac myocytesfusion cellsmyocardial infarction
0
浏览量
0
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621