查找使用阿拉丁产品的科研文献

A Combined Model of Human iPSC-Derived Liver Organoids and Hepatocytes Reveals Ferroptosis in DGUOK Mutant mtDNA Depletion Syndrome

Jingyi Guo,Lifan Duan,Xueying He,Shengbiao Li,Yi Wu,Ge Xiang,Feixiang Bao,Liang Yang,Hongyan Shi,Mi Gao,Lingjun Zheng,Huili Hu,Xingguo Liu

Mitochondrial DNA depletion syndrome (MDS) is a group of severe inherited disorders caused by mutations in genes, such as deoxyribonucleoside kinase (DGUOK). A great majority of DGUOK mutant MDS patients develop iron overload progressing to severe liver failure. However, the pathological mechanisms connecting iron overload and hepatic damage remains uncovered. Here, two patients’ skin fibroblasts are reprogrammed to induced pluripotent stem cells (iPSCs) and then corrected by CRISPR/Cas9. Patient-specific iPSCs and corrected iPSCs-derived high purity hepatocyte organoids (iHep-Orgs) and hepatocyte-like cells (iHep) are generated as cellular models for studying hepatic pathology. DGUOK mutant iHep and iHep-Orgs, but not control and corrected one, are more sensitive to iron overload-induced ferroptosis, which can be rescued by N-Acetylcysteine (NAC). Mechanically, this ferroptosis is a process mediated by nuclear receptor co-activator 4 (NCOA4)-dependent degradation of ferritin in lysosome and cellular labile iron release. This study reveals the underlying pathological mechanisms and the viable therapeutic strategies of this syndrome, and is the first pure iHep-Orgs model in hereditary liver diseases.

Noninvasive Optogenetics Realized by iPSC-Derived Tentacled Carrier in Alzheimer's Disease Treatment

Yuewen Zhai,Fan Gao,Shihao Shi,Qifeng Zhong,Jinnan Zhang,Ji Fang,Fang He,Yanqin Zhang,Yu Li,Fei Liu,Bing Xue,Yueqing Gu,Siwen Li

Neural-activated optogenetics technique contributing to the “restart” of degenerative neurons offers hope for the treatment of several neurodegenerative diseases. However, the limitations of persistent invasiveness and inadequate repair of the pathological environment strongly hinder its further application. Here, a concept of differentiating stem cells is proposed to produce functional materials to enhance the therapeutic applicability of optogenetics. Induced pluripotent stem cells (iPSCs) are differentiated to generate the “tentacled” stem cells TenSCs. Their “tentacled” vesicles TenSCev, upon inheriting the biological functions of the parent cell, will possess both neural targeting capacity and pathological environment repair ability. Hence, TenSCev are utilized as functional carrier to deliver optogenetics elements for completely non-traumatic and controllable neuron activation, while also facilitating the comprehensive restoration of the pathological environment, thus effectively halted disease progression and significantly improved cognitive function in Alzheimer's disease or aged mice. Further, the concept of generating specialized biomaterials from differentiated stem cells as functional carriers holds the potential to broaden the applicability of various neuroregulatory techniques in the treatment of neurological disorders.

BRG1 enhances porcine iPSC pluripotency through WNT/β-catenin and autophagy pathways

Xuan Ren,Jianchun Xu,Qingsong Xue,Yi Tong,Tairan Xu,Jinli Wang,Ting Yang,Yuan Chen,Deshun Shi,Xiangping Li

Brahma-related gene 1 (BRG1) enhances the pluripotency of embryonic and adult stem cells , however, its effect on induced pluripotent stem cell (iPSC) pluripotency has not been reported. The aim of this study was to investigate the effect of BRG1 on porcine iPSC pluripotency and its mechanisms. The effect of BRG1 on porcine iPSC pluripotency was explored by positive and negative control it. The mechanism was investigated by regulating the WNT/β-catenin signaling pathway and autophagy flux. The results showed that inhibition of BRG1 decreased pluripotency-related gene expression in porcine iPSCs; while its overexpression had the opposite effect, the expression of WNT/β-catenin signaling pathway- and autophagy-related genes was significantly up-regulated ( P  < 0.05) in the BRG1 overexpressed group when compared to the control group. Inhibited pluripotency-related gene or protein expression , decreased autophagy flux, and increased mitochondrial length and mitochondrial membrane potential (MMP) were observed when porcine iPSCs were treated with the WNT/β-catenin signaling pathway inhibitor IWR-1. Forced BRG1 expression restored porcine iPSC pluripotency, increased autophagy flux, shortened mitochondria, and reduced MMP. Lastly, Compound C was used to activate porcine iPSC autophagy, and it was found that the expression of BRG1 and β-catenin increased, and pluripotency-related gene and protein expression was up-regulated; these effects were reversed when the BRG1 inhibitor PFI-3 and IWR-1 were added. These results suggested that BRG1 enhanced the pluripotency of porcine iPSCs through WNT/β-catenin and autophagy pathways.

Effect of Honokiol on culture time and survival of Alzheimer’s disease iPSC-derived neurons

Duong Thi Thuy Le,Cuong Manh Vu,Thuy Thi Bich Ly,Nam Trung Nguyen,Phuong Thi Mai Nguyen,Ha Hoang Chu

Medium from human iPSC-derived primitive macrophages promotes adult cardiomyocyte proliferation and cardiac regeneration

Xiao Yi,Zhang Hao,Liu Xu,Xu Pengfei,Du Heng,Wang Jiawan,Shen Jianghua,Li Yujing,Wang Yuhan,He Chuting,Feng Haiping,Liu Jingfang,Zhou Yanan,Liu Siqi,Gao Zeyu,Zang Jingyi,Bi Jinmiao,Tang Tie-Shan,Gu Qi,Wei Tuo,Wang Jun,Song Moshi

Heart injury has been characterized by the irreversible loss of cardiomyocytes comprising the contractile tissues of the heart and thus strategies enabling adult cardiomyocyte proliferation are highly desired for treating various heart diseases. Here, we test the ability of human induced pluripotent stem cell-derived primitive macrophages (hiPMs) and their conditioned medium (hiPM-cm) to promote human cardiomyocyte proliferation and enhance cardiac regeneration in adult mice. We find that hiPMs promote human cardiomyocyte proliferation, which is recapitulated by hiPM-cm through the activation of multiple pro-proliferative pathways, and a secreted proteome analysis identifies five proteins participating in this activation. Subsequent in vivo experiments show that hiPM-cm promotes adult cardiomyocyte proliferation in mice. Lastly, hiPM-cm enhances cardiac regeneration and improves contractile function in injured adult mouse hearts. Together, our study demonstrates the efficacy of using hiPM-cm in promoting adult cardiomyocyte proliferation and cardiac regeneration to serve as an innovative treatment for heart disease.

Magnetically Aligned Fiber-Reinforced ECM Patch Loaded With iPSC-CMs Improves Cardiac Repair After Myocardial Infarction

Xinming Wang,Weichang Xie,Zichen Wang,Haonan Zhang,Jia Li,Xingjian Gao,Dingjie Zhou,Huaiyu Jiang,Shixing Huang,Qiang Zhao,Xiaofeng Ye

Repairing anisotropic tissues such as the myocardium requires implanted cells to organize into highly ordered structures. However, achieving controlled cell alignment remains a major challenge for current cell delivery patches. Here, we develop a magnetically aligned, fiber-reinforced extracellular matrix (ECM) patch that directs anisotropic cell organization and supports stem cell implantation. Poly-lactic acid fibers containing Fe 3 O 4 are fabricated via electrospinning and controlled hydrolysis, and are aligned uniaxially within ECM hydrogels using an external magnetic field prior to gelation. The resulting ECM–fiber patches exhibit enhanced mechanical strength and reliably guide the alignment of cardiomyocytes, fibroblasts, and endothelial cells. A beating patch is also created by seeding human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). In a nude mouse myocardial infarction (MI) model, implantation of iPSC-CMs@ECM–fiber patches preserves cardiac function, reduces apoptosis, limits fibrosis, and enhances vascularization. Implanted iPSC-CMs demonstrate high viability and low immunogenicity. Molecular profiling reveals transient modulation of early inflammatory responses post-MI, probably mediated through IL-17, IL-3, and IL-1β pathways. Overall, the ECM–fiber patch provides a versatile platform for delivering highly organized cells for in vivo tissue repair.

A NANOG-EGFP iPSC-Based Reporter Platform for High-Throughput Screening of Environmental Pollutant Effects on Pluripotency

Yanyi Zhao,Hanyue Li,Renjun Yang,Nuoya Yin,Francesco Faiola

Humans are frequently exposed to diverse environmental pollutants that may pose potential risks during early embryonic development. Advances in stem cell toxicology have introduced new opportunities for environmental toxicity assessment, enabling the development of physiologically relevant models based on human induced pluripotent stem cells (iPSCs). However, current systems remain limited by low throughput and the inability to achieve real-time monitoring. In this study, we established a NANOG-EGFP iPSC reporter line by precisely inserting a T2A-EGFP cassette into the endogenous NANOG locus using CRISPR/Cas9, enabling dynamic and quantitative visualization of pluripotency status. Using this reporter system, we systematically evaluated the effects of 38 representative environmental pollutants spanning six major chemical categories, including bisphenols, per- and polyfluoroalkyl substances (PFAS), halogenated flame retardants, organophosphate esters, neonicotinoids, and phenolic compounds. Most pollutants induced varying degrees of reduction in NANOG-EGFP expression, suggesting perturbation of core transcriptional networks involved in pluripotency maintenance rather than nonspecific cytotoxic effects. Notably, bisphenol B (BPB), bisphenol Z (BPZ), perfluorobutanesulfonic acid (PFBS), tributyl phosphate (TnBP), tris(2-chloroethyl) phosphate (TCEP), and 3,5-di-tert-butyl-4-hydroxybenzoic acid (BHT-COOH) exhibited pronounced low-dose activity, causing significant suppression of NANOG-EGFP signals even at 10 nmol/L. Furthermore, during neuroectoderm differentiation, several bisphenols and PFAS delayed the normal downregulation of NANOG, indicating disruption of the tightly regulated timing of pluripotency exit and lineage transition during early developmental progression. Overall, the NANOG-EGFP iPSC reporter model provides a sensitive, stable, and practical platform for high-throughput screening of environmental toxicants and offers mechanistically informed insight into how pollutant exposure may interfere with early human developmental regulatory programs.

3D-Printed Collagen/Silk Fibroin/Matrigel Scaffold Combined with iPSC-Derived Cells for Tissue Regeneration of Salivary Glands

Liu Han,Sun Ziyan,Jin Yizhou,Li Ruixin,Shan Zhaochen

Salivary gland defects from tumors or trauma are often accompanied by irreversible secretory dysfunction and remain a clinical challenge. Alternative cell sources and suitable scaffolds for salivary gland regeneration remain lacking. A functionalized scaffold system is fabricated by three-dimensional printing combined with freeze-drying to prepare a collagen-silk fibroin-Matrigel scaffold. The scaffold exhibits high porosity and strong water absorption capacity. Its internal surface is coated with a basement membrane extract-rich layer that enhances hydrophilicity and creates a physiologically relevant cell niche. In vitro, a synergistic co-culture system incorporating urine-derived induced pluripotent stem cells (iPSCs), parotid mesenchymal stem cells, and parotid epithelial cells directs the differentiation of iPSCs into KRT18 + /VIM + /AQP5 + iPSC-derived cells. These cells readily adhere to the scaffold, proliferate, and form acinar-like aggregates within its porous architecture. In a rat model of parotid gland trauma, implantation of the scaffold progressively promotes tissue regeneration and attenuates trauma-induced atrophy. Histological analysis reveals that the scaffold recruits host cells to organize within its pores, forming amylase-secreting acinar-like structures. This study provides a reliable seed cell source and a suitable scaffold for structural and functional parotid restoration, offering a potential strategy for salivary gland tissue engineering.

A composite hydrogel improves the survival and differentiation of human iPSC-derived neural stem cells after ischemic stroke

Yang Liu,Wen Zhang,Cheng Hu,Cheng Zheng,Fanjun Zhang,Li Yang,Zhengyong Li,Yunbing Wang

Stem cell transplantation can replace lost and damaged neurons after ischemic stroke and has been proposed as a potential treatment for stroke. However, stem cell therapy is challenged by poor cell retention, survival, and differentiation following transplantation. In this study, we developed a composite hydrogel incorporating brain decellularized extracellular matrix (dECM) and a matrix metallopeptidase (MMP)-responsive host–guest (HG) hydrogel that acts as a platform for human induced pluripotent stem cell-derived neural stem cells (iPSC-NSCs) delivery to improve transplant stem cell survival and differentiation. The optimized composite hydrogel ratio was screened by rheology analysis and neurite outgrowth assessment. We then examined the effects of HG/dECM composite hydrogels on iPSC-NSCs survival and differentiation on two-dimensional surfaces modified with hydrogel and in a three-dimensional hydrogel network. Moreover, iPSC-NSCs-laden composite hydrogels were injected into the infarct cavity in an ischemic stroke model to replace lost and damaged nerve cells. As expected, the HG/dECM composite hydrogels better facilitated the survival and differentiation of iPSC-NSC in vivo than the control. Likewise, the iPSC-NSCs loaded in HG/dECM composite hydrogels showed superior retention and survival in the cortical infarct cavity, and the composite hydrogels promoted the differentiation of iPSC-NSCs into neurons. Overall, HG/dECM composite hydrogels are a promising platform to regulate stem cell survival and differentiation.

A PEDOT:PSS-MWCNT-modified MEA platform with integrated electrical stimulation for enhanced maturation and drug screening of iPSC-cardiomyocytes

Zhixiang Liang,Huimin Li,Junlei Han,Jun Chen,Jiemeng Ding,Wenteng Tang,Minghui Wang,Qi Meng,Lei Zhang,Zetao Zhang,Li Wang

Cardiovascular toxicity is a key safety indicator that has to be evaluated in the process of new drug development. To enable high-throughput and multiparametric assessment of cardiomyocyte function, we developed a 32-well high-throughput microelectrode array (MEA) platform integrated with electrical stimulation. Electrode surface modification with PEDOT:PSS-MWCNT nanocomposites significantly enhanced electrochemical performance and signal acquisition fidelity, reducing electrode impedance by 37-fold compared to unmodified Au electrodes. Notably, the built-in electrical stimulation effectively promoted electrophysiological and structural maturation of human iPSC-cardiomyocytes, demonstrated by a 15.47% increase in sarcomere length and 1.73-fold upregulation of connexin-43 expression. The platform's utility was further validated through cardiotoxicity assessment of two representative ion channel modulators, E-4031 (a selective hERG channel blocker) and verapamil (a clinically used calcium channel blocker). Collectively, this study presents an integrated platform that combines functional monitoring with cellular maturation enhancement, offering a powerful tool for cardiotoxicity screening and disease modeling.

Using low-risk factors to generate non-integrated human induced pluripotent stem cells from urine-derived cells

Wang Linli,Chen Yuehua,Guan Chunyan,Zhao Zhiju,Li Qiang,Yang Jianguo,Mo Jian,Wang Bin,Wu Wei,Yang Xiaohui,Song Libing,Li Jun

Background Because the lack of an induced pluripotent stem cell (iPSC) induction system with optimal safety and efficiency limits the application of these cells, development of such a system is important.Methods To create such an induction system, we screened a variety of reprogrammed plasmid combinations and multiple compounds and then verified the system’s feasibility using urine cells from different individuals. We also compared large-scale iPSC chromosomal variations and expression of genes associated with genomic stability between this system and the traditional episomal system using karyotype and quantitative reverse transcription polymerase chain reaction analyses.Results We developed a high-efficiency episomal system, the 6F/BM1-4C system, lacking tumorigenic factors for human urine-derived cell (hUC) reprogramming. This system includes six low-risk factors (6F), Oct4, Glis1, Klf4, Sox2, L-Myc, and the miR-302 cluster. Transfected hUCs were treated with four compounds (4C), inhibitor of lysine-demethylase1, methyl ethyl ketone, glycogen synthase kinase 3 beta, and histone deacetylase, within a short time period. Comparative analysis revealed significantly decreased chromosomal variation in iPSCs and significantly increased Sirt1 expression compared with iPSCs induced using the traditional episomal system.Conclusion The 6F/BM1-4C system effectively induces reprogramming of urine cells in samples obtained from different individuals. iPSCs induced using the 6F/BM1-4C system are more stable at the cytogenetic level and have potential value for clinical application.

Human iPSC-derived mesenchymal stem cells relieve high blood pressure in spontaneously hypertensive rats via splenic nerve activated choline acetyltransferase-positive cells

Zhang Zhen,Huang Weijun,Zhang Xiaoran,Wang Zhecun,Xie Manting,Xie Bingbing,Wang Yiling,Chen Xiaoyong,Xiang Andy Peng,Xiang Qiuling

Despite substantial advancements in modern medicine, the management of hypertension remains a major challenge. Stem cell-based therapies have recently demonstrated remarkable efficacy in treating cardiovascular diseases, including hypertension. However, the antihypertensive mechanism of mesenchymal stem cells (MSCs) has not been extensively explored. This study aimed to investigate the role of injected MSCs in regulating blood pressure homeostasis. Our previous study demonstrated that human induced pluripotent stem cell-derived mesenchymal stem cells (hiPSC-MSCs) are functional and homogeneous sources for MSC-based therapy. After the injection of hiPSC-MSCs, a significant reduction in blood pressure and end target organ inflammation were observed in spontaneously hypertensive rats (SHRs). Cell tracking assays demonstrated that the injected hiPSC-MSCs accumulated in the spleens of the SHRs. The injected hiPSC-MSCs accumulated adjacent to the splenic nerve, potentially contributing to the antihypertensive effects. Furthermore, the hiPSC-MSCs released abundant glutamate, which acts as a neuromodulator to activate the splenic sympathetic nerve. After inhibition of glutamate synthesis by siRNA, the ability of hiPSC-MSCs to activate sympathetic nerves was significantly diminished. In addition, the antihypertensive effects of hiPSC-MSCs were eliminated after splenic nerve denervation (SND), underscoring the critical role of the splenic nerve. Moreover, activation of the splenic nerve resulted in increased release of norepinephrine (NE), which increased the number of choline acetyltransferase-positive (ChAT + ) cells in the spleen and peripheral blood. Consequently, the acetylcholine (ACh) produced by elevated ChAT + cells could act as a vasodilator, lowering blood pressure and mitigating inflammation in end target organs. In summary, our findings indicate that hiPSC-MSCs effectively lower blood pressure in hypertension by influencing the splenic nerves and regulating ChAT + cells. The connection between blood pressure regulation and the splenic nerve may offer new insights into the treatment of hypertension.