Genetic modification of primate amniotic fluid-derived stem cells produces pancreatic progenitor cells in vitro.
Genetic modification of primate amniotic fluid-derived stem cells produces pancreatic progenitor cells in vitro.
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DOI：
10.1159/000345816
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Soker S
中科院分区：
文献类型：
--
作者：
Zhou Y;Mack DL;Williams JK;Mirmalek-Sani SH;Moorefield E;Chun SY;Wang J;Lorenzetti D;Furth M;Atala A;Soker S
1型糖尿病（T1D）的胰岛素治疗不能预防严重的长期并发症，包括血管疾病、神经病变、视网膜病变和肾功能衰竭。干细胞，包括羊水衍生干细胞（AFS）细胞——高度膨胀、多能、非致瘤性细胞——可以作为β细胞分化的合适干细胞来源。在目前的研究中，我们测试了非人灵长类动物（nhp）的AFS细胞是否能够在体外分化为β样细胞表型。对CD117 （c-kit）阳性群体采用免疫磁选法从食蟹猴羊水中获得nhpas细胞。在腺病毒介导了PDX1、NGN3和MAFA的表达后，对AFS细胞进行了内胚层和胰腺谱系特异性标志物的RT-PCR检测。在nhpaafs细胞系中，MAFA的表达足以诱导胰岛素mRNA的表达，而MAFA与PDX1和ngn3的联合作用进一步诱导胰岛素的表达，并诱导其他重要内分泌细胞基因如胰高血糖素、NEUROD1、NKX2.2、ISL1和PCSK2的表达。通过在添加B27、β细胞素和烟酰胺的培养基中培养三重感染的AFS细胞，以及在细胞外基质包被板上培养细胞，可以获得更高的这些和其他重要胰腺基因的诱导。胰腺基因如NEUROD1、胰高血糖素和胰岛素的表达随着腺病毒表达的PDX1、NGN3和MAFA的下降而逐渐降低。总之，这些实验表明，灵长类动物AFS细胞中胰腺转录因子的强制表达诱导它们向胰腺谱系发展。
Insulin therapy for Type 1 diabetes (T1D) does not prevent serious long-term complications including vascular disease, neuropathy, retinopathy and renal failure. Stem cells, including amniotic fluid-derived stem (AFS) cells--highly expansive, multipotent, and non-tumorigenic cells--could serve as an appropriate stem cell source for β-cell differentiation. In the current study we tested whether nonhuman primate (nhp) AFS cells ectopically expressing key pancreatic transcription factors were capable of differentiating into a beta-like cell phenotype in vitro. NHPAFS cells were obtained from Cynomolgus monkey amniotic fluid by immunomagnetic selection for a CD117 (c-kit) positive population. RT-PCR for endodermal and pancreatic lineage-specific markers was performed on AFS cells after adenovirally transduced expression of PDX1, NGN3 and MAFA. Expression of MAFA was sufficient to induce insulin mRNA expression in nhpAFS cell lines, whereas a combination of MAFA, PDX1 and NGN3further induced insulin expression, as well as induced the expression of other important endocrine cell genes such as glucagon, NEUROD1, NKX2.2, ISL1 and PCSK2. Higher induction of these and other important pancreatic genes was achieved by growing the triply infected AFS cells in media supplemented with a combination of B27, betacellulin and nicotinamide, as well as culturing the cells on extra-cellular matrix coated plates. The expression of pancreatic genes such as NEUROD1, glucagon and insulin progressively decreased with the decline of adenovirally-expressed PDX1, NGN3 and MAFA. Together, these experiments suggest that forced expression of pancreatic transcription factors in primate AFS cells induces them towards the pancreatic lineage.
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影响因子：
2.9
作者：
Hui, Hongxiang;Tang, Yongming G.;Go, Vay Liang W.
通讯作者：
Go, Vay Liang W.
影响因子：
5.2
作者：
Jiang, Jianjie;Au, Melinda;Majumdar, Anish S.
通讯作者：
Majumdar, Anish S.
影响因子：
23.9
作者：
Borowiak M;Maehr R;Chen S;Chen AE;Tang W;Fox JL;Schreiber SL;Melton DA
通讯作者：
Melton DA
DOI：
10.1111/j.1582-4934.2010.01034.x
发表时间：
2011-03-01
影响因子：
5.3
作者：
Chiou, Shih-Hwa;Chen, Shih-Jen;Ku, Hung-Hai
通讯作者：
Ku, Hung-Hai
影响因子：
5.4
作者：
De Carlo, E.;Baiguera, S.;Parnigotto, P. P.
通讯作者：
Parnigotto, P. P.