人多能干細胞研究  目錄

hiPS細胞建系

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為您實現將疾病模型培養在培養皿中的理想

Generation of Integration-Free iPS Cells From 5ml Human Blood

如何取的合適的樣本？

如何防止樣本被污染？

實驗室實驗條件是否能夠滿足？

如何高效率重編程生成hiPS細胞？

          。。。。。。。。。。？？？

還在為取材困難而無法建立疾病模型煩惱嗎？我們可利用Episomal  iPSC Reprogramming高效率重編程系統將5-10 mL外周血經轉染后即可誘導生成無外源基因插入整合的hiPSCs，為您輕松建立理想的疾病模型，用于iPSCs疾病模型的建立、藥物開發、細胞治療以及其它再生醫學研究領域。

適合應用領域：

1. iPS疾病模型的建立

2. 創傷樣本無法取材

3. 疾病模型用于藥物開發

4. 細胞治療以及再生醫學研究

5. 尿液樣本量少，無法滿足實驗需求

hiPSCs細胞鑒定：

載體信息：

pEV-SFFV-OCT4-Wpre; 

pEV-SFFV-SOX2-Wpre; 

pEV-SFFV-Myc-Wpre; 

pEV-SFFV-KLF4-Wpre; 

pEV-SFFV-OCT4-E2A-SOX2-Wpre;

pEV-SFFV-MYC-E2A-KLF4-Wpre; 

pEV-SFFV-BCL-XL-Wpre

注：

SFFV: spleen focus-forming virus U3 promoter; 

 E2A: a self-cleavage site derived from equine rhinitis A virus,(CAG TGT ACT AAT TAT GCT CTC TTG AAA TTG GCT GGA GAT GTT GAG AGC AAC CCA GGT CCC); 

WPRE: posttranscriptional regulatory element; Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) is a DNA sequence that, when transcribed, creates a tertiary structure enhancing expression. The sequence is commonly used in molecular biology to increase expression of genes delivered by viral vectors.

SV40PolyA: polyadenylation signal from SV40 virus; 

OriP: EBV origin of replication; 

EBNA1: Epstein–Barr nuclear antigen 1, which plays essential roles in replication and persistence of episomal plasmid in infected cells.

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參考文獻：

1. Meng X, Neises A, Su RJ, Payne KJ, Ritter L, Gridley DS, Wang J, Sheng M, Lau KH, Baylink DJ, Zhang XB: Efficient reprogramming of human cord blood CD34+ cells into induced pluripotent stem cells with OCT4 and SOX2 alone. Molecular Therapy, 2012;20(2):408-16.

2. Su RJ, Baylink DJ, Neises A, Kiroyan JB, Meng X, Payne KJ, Tschudy-Seney B, Duan Y, Appleby N, Kearns-Jonker M, Gridley DS, Wang J, Lau KHW, Zhang XB: Efficient generation of integration-free iPS cells from human adult peripheral blood using BCL-XL together with Yamanaka factors. PLoS One. 2013, 8(5): e64496.

3. Zhang XB: Cellular reprogramming of human peripheral blood cells. Genomics Proteomics Bioinformatics. 2013 Oct;11(5):264-74.

4. Su RJ, Neises A, Zhang XB: Generation of iPS cells from human peripheral blood mononuclear cells using episomal vectors. Methods Mol Biol. 2016;1357:57-69.

5. Wen W#, Zhang JP#, Xu J#, Su RJ, Neises A, Ji GZ, Yuan W, Cheng T*, Zhang XB*. Enhanced generation of integration-free iPSCs from human adult peripheral blood mononuclear cells with an optimal combination of episomal vectors. Stem Cell Reports. 2016 Jun 14;6(6):873-84.

6. Wen W, Zhang JP, Chen W, Arakaki C, Li XL, Baylink D, Botimer GD, Xu J, Yuan W, Cheng T*, Zhang XB*. Generation of integration-free induced pluripotent stem cells from human peripheral blood mononuclear cells using episomal vectors. J Vis Exp. 2017 Jan 1;(119). doi: 10.3791/55091.

 人多能干細胞研究  目錄