International journal of stem cells最新文献

Ovarian cancer remains one of the most lethal gynecologic malignancies, with limited responsiveness to standard chemotherapy and poor long-term prognosis. Tumor-associated macrophages, particularly M2-polarized populations, play a crucial role in immune suppression and tumor progression. Human induced pluripotent stem cells (hiPSCs) can differentiate into functional immune cells, providing an unlimited and patient-specific source for cell-based immunotherapy. In this study, we investigated the therapeutic potential of hiPSC-derived macrophages (hiMACs), specifically M1-polarized hiMACs, against ovarian cancer. In a co-culture system, M1-hiMACs significantly reduced the viability of ovarian cancer cells, inducing apoptosis and necrosis, whereas M0 macrophages showed minimal effects. In vivo, intravenous administration of M1-hiMACs into nude mice bearing ovarian cancer cells resulted in a dose-dependent reduction in tumor volume. Furthermore, combination therapy with paclitaxel and M1-hiMACs led to greater tumor regression and enhanced histological necrosis compared to either treatment alone. These findings demonstrate the potent anti-tumor effects of M1-hiMACs and highlight their potential for cellular immunotherapy for ovarian cancer, particularly in combination with chemotherapy.

Human pluripotent stem cells (hPSCs) can self-renew indefinitely and differentiate into all three germ layers. However, the primary regulators of hPSC cell cycle dynamics remain unclear. To identify novel regulators of hPSC proliferation, transcriptomic profiling of undifferentiated hPSCs and somatic cells was performed via next-generation sequencing. Stomatin-like protein 2 (STOML2) and prohibitin (PHB) were among the upregulated genes in hPSCs and were closely associated with the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling pathway. Temporal expression analysis indicated that STOML2 and PHB decreased during differentiation and increased during reprogramming. Short hairpin RNA (shRNA)-mediated knockdown of STOML2 in hPSCs caused phenotypic changes. Gene expression analyses demonstrated reduced OCT4, NANOG, PHB, and phosphorylated ERK, alongside increased differentiation markers across all three germ layers. The STOML2-PHB axis is essential for maintaining hPSC identity by sustaining ERK/MAPK activity and cell cycle structure. This study identified STOML2 as a key pluripotency regulator and provides new insight into intrinsic stem cell fate control.

Cardiotoxicity assessment is a crucial step in the drug development process. With growing interest in alternatives to animal testing, preclinical cardiotoxicity evaluation has become increasingly important. Human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) offer a physiologically relevant in vitro model for this purpose. As a result, electrophysiological analysis platforms using iPSC-CMs have gained attention. However, conventional microelectrode array (MEA) chips rely on metal electrodes, which are costly and optically opaque. This lack of transparency limits detailed morphological observation of the cells. In this study, we employed an MEA chip incorporating transparent and conductive indium tin oxide (ITO) electrodes to simultaneously monitor both morphological changes and field potential (FP) of iPSC-CMs. iPSC-CMs cultured on ITO chips exhibited stable electrophysiological signals reflecting coupled depolarization and repolarization, along with self-organization. Short-term exposure to ion channel blockers did not induce noticeable morphological alterations; however, dose-dependent changes in FPs were observed. In contrast, treatment with cardiotoxic drugs resulted in morphological damage and reduced cell viability, accompanied by progressive alterations in key FP parameters over the treatment period. These findings demonstrate the potential of ITO-based MEA as next-generation cardiotoxicity evaluation platforms capable of real-time monitoring of both drug-induced electrophysiological responses and optical cellular changes.

Pyrene (Pyr), a representative subtype of polycyclic aromatic hydrocarbons, is primarily generated during the incomplete combustion of organic matter. As an environmental pollutant, Pyr has been reported to exert adverse effects on various physiological systems. However, data regarding its cardiotoxicity remain limited. In this study, we investigated the acute cardiotoxic effects of Pyr using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). hiPSC-CMs were exposed to various doses of Pyr, and both cell viability and functional parameters were evaluated. Pyr did not affect cell viability under 30-minute and 2-hour exposure conditions regardless of dose. However, significant differences were observed in the dual-cardiotoxicity evaluation based on a microelectrode array, which allows simultaneous assessment of electrophysiological signals and contractility in CMs. Pyr decreased beat period and field potential duration in a dose-dependent manner, resembling the acute cardiotoxicity pattern of I_Kr and I_CaL channel blockers, and progressively reduced spike amplitude over time. Pyr also progressively reduced spike amplitude over time. Although a transient decrease in beat amplitude was observed at high dose, it recovered over time, while the excitation-contraction delay was reduced. Taken together, these findings demonstrate that Pyr can induce functional cardiotoxicity even under acute exposure and highlight the value of the established evaluation method for the development of safer alternative substances.

A rare disease is generally defined as a medical condition that affects a small proportion of the population, though specific thresholds vary across countries. Despite regional differences, these definitions consistently reflect the low prevalence of such conditions, the limited availability of effective treatments, and the pressing need for targeted research and regulatory support. As a result of their rarity and low commercial potential, rare diseases have historically represented an area of market failure, where investment and research have been limited and often neglected. However, since the 1990s, each country has guaranteed continuous support to research and development projects to promote the advancements of rare disease treatments, achieving a growth rate greater than that of the entire pharmaceutical industry. In this review, we examine the status of orphan drug development using an advanced therapy medicinal product (ATMP) approach in the growing rare disease market, with a particular focus on cell therapies and gene therapies, which constitute the most actively developed and clinically applied categories within ATMPs. We also explore strategic approaches through which the orphan drug industry can utilize ATMPs, especially these two modalities, to enhance its competitiveness.

Bronchopulmonary dysplasia (BPD), characterized by impaired alveolarization and dysregulated vascularization, is a severe health burden for neonates worldwide. Hyperoxia induced acute lung injury is a major contributor to the progression and deterioration of BPD. An increasing number of animal studies have revealed that human umbilical cord blood derived mononuclear cells (hUCB-MNCs) infusion significantly attenuated the hyperoxia-induced acute lung injury through regeneration capacity. Currently, clinical application requires determination of the optimal dose and adjustment to good manufacturing practices. In this work, we comprehensively investigated the optimal dose of hUCB-MNCs in alleviating hyperoxia-induced lung injury in neonatal C57BL6/J mice. Mice with hyperoxia exposure were implanted with low (3×10⁴ cells/kg, Dl), middle (3×10⁵ cells/kg, Dm) and high (3×10⁶ cells/kg, Dh) dose of hUCB-MNCs at postnatal day 7. Three weeks after graft, characteristics exhibited in lungs including morphology, function and cytokine expression were thoroughly analyzed. Implantation of hUCB-MNCs sharply reverted the impaired lung architecture induced by hyperoxia exposure dose dependently as evidenced by indicated parameters. Attenuated expression of IL-1β concomitant with enhanced expression of IL-10 and IL-2 were shown in Dh inoculated groups, where Dl and Dm failed to restore the level of IL-10, IL-1β and IL-2. Significantly re-escalated marker of angiogenic marker VEGFA, CD31 in lung tissue were uniquely observed in Dh group. Mechanistically, our study revealed the appropriate dose of intravenous infusion of hUCB-MNCs in alleviating hyperoxia-induced lung injury through modulating reactive oxygen species response in neonatal mice. Therefore, a tight control of hUCB-MNCs density or levels of CB-MNC related products is of great significance.

There is mounting epidemiologic and experimental evidence of the harmful effects of exposure to fine particulate matter (PM2.5) on human lung health. However, the current utilization of in vitro two-dimensional (2D) cell culture and in vivo animal models falls short in accurately recapitulating the complexity, functions, and development of the human lung. Recently, technologies for creating 3D biomimetic lung organoids from human tissue and pluripotent stem cells that mimic the structure and function of human organs have been rapidly developed. Human lung organoids are currently being applied in various studies such as disease modeling, drug efficacy testing, and regenerative medicine. In the field of environmental toxicology, human lung organoids offer a promising solution for more accurately assessing the health impacts of exposure to PM2.5 and to the limitations posed by 2D cultures and animal models. In this review, we summarize the effects of PM2.5 exposure on human lung and other organoids, as well as the molecular and genetic impacts of such exposure using human organoids. Furthermore, the development of lung-on-a-chip technology to mimic the microenvironment and its utilization for evaluating the pulmotoxicity of PM2.5 exposure are discussed.

Human pluripotent stem cells (hPSCs) can be used to investigate hematopoietic development and have the potential to advance cell-based therapies and to facilitate developmental biology studies. However, efficient ex vivo differentiation into hematopoietic lineages, including red blood cells (RBCs) of the erythroid lineage and immune cells such as macrophages of the myeloid lineage, is hampered by the need for precise temporal regulation of cytokines and growth factors. In this study, we developed an optimized protocol for hematopoietic lineage specification from hPSCs by fine-tuning the temporal dynamics of cytokine and growth factor applications. Prolonged mesodermal specification in the absence of hemogenic cytokines significantly enhanced the generation of hematopoietic progenitors (CD34^＋CD45^＋) with robust functional potential. Early administration of interleukin (IL)-3 during hematopoietic specification promoted progenitor expansion and maturation. Supplementation of bone morphogenetic protein 4 at the hematopoietic maturation stage enhanced the differentiation efficiency and preferentially drove myeloid lineage commitment toward macrophages at the expense of erythroid differentiation. The timing of erythropoietin administration was important in erythroid lineage commitment, and delayed treatment (day 10) enhanced erythroblast expansion and RBC production. By contrast, the timing of IL-6, GM-CSF, and M-CSF exposure did not significantly affect macrophage differentiation efficiency, suggesting that myeloid lineage specification follows a default pathway under optimized differentiation conditions. These findings suggest a refined, time-controlled strategy for directing hematopoietic differentiation from hPSCs, and provide insight into therapeutic blood cell production, regenerative medicine, and ex vivo modeling of hematopoietic disorders.

Phenylketonuria (PKU), an autosomal recessive genetic disorder, has been documented to exhibit over 950 distinct mutations. This condition primarily affects the metabolism of phenylalanine, which is affected by a deficiency in the hepatic enzyme phenylalanine hydroxylase. The optimal treatment for PKU disease remains to be determined, necessitating further research. The severity of the disease and the most effective treatment method vary depending on the specific mutation, which necessitates the development of personalized treatment strategies. In this study, we successfully established induced pluripotent stem cell (iPSC) lines from the blood of a PKU patient with the R243Q mutation via Sendai virus-based reprogramming (R243Q-iPSCs). The established R243Q-iPSCs exhibited characteristics of pluripotency, as confirmed through quantitative reverse transcription polymerase chain reaction, western blot, immunocytochemistry, and karyotype analysis. Furthermore, these iPSCs not only successfully differentiated into hepatocytes but also exhibited a complete PKU disease phenotype. These results provide a valuable foundation for PKU disease research, including physiological studies of PKU, gene therapy, drug screening, and the development of platforms for novel cell therapy approaches.

Nanog is a key transcription factor that regulates the self-renewal and pluripotency of embryonic stem cells (ESCs). Although Kap1 has been demonstrated to regulate the stability of stemness factors, including Oct4 and Lin28A, its role in regulating Nanog protein stability in ESCs remains unexplored. In the present study, we examined the interaction between Kap1 and Nanog and its role in stabilizing the Nanog protein. Immunoprecipitation assays revealed that Nanog specifically interacted with the coiled-coil domain of Kap1. Kap1 overexpression increased the stability of the Nanog protein by inhibiting its ubiquitination and proteasomal degradation, whereas Kap1 silencing accelerated Nanog degradation. Furthermore, Kap1 overexpression inhibits Nanog degradation by interfering with the binding of Nanog to Fbxw8, an E3 ubiquitin ligase that promotes Nanog degradation via a proteasome-dependent process. These results indicate that Kap1 acts as a key regulator to preserve ESC properties by modulating the protein stability of stemness factors, including Oct4, Lin28A, and Nanog.