Cells Tissues Organs最新文献

Introduction: Cellular wound healing assay is an important experimental technique for detecting cell migration in vitro. Scratching on monolayer cells using a pipette tip is commonly used. However, it is difficult to guarantee the scratch with the same width, and the initial scratch width has a large impact on the experimental results for different treatment factors or different cell types. To optimize this assay for diverse experimental requirements, we developed an experimental device capable of generating scratches with variable widths.

Methods: Our device offers the flexibility of selecting among four widths to create cell scratches, enabling the choice of an optimal initial scratch width for specific cell types and experimental conditions.

Results: This device produced straight, clean wounds with precise widths. Comparing cell growth in the four width wounds, Hepa1-6 and HUMSCs showed the greatest difference in 0.6 cm wound, 143B at 0.9 cm wound and urine-derived stem cells at 1.2 cm wound were significantly different, which suggests that the width of the wounds has a huge impact on the experimental results. Compared to other wound inserts on the market, our device is more efficient and economical.

Conclusion: This versatile and practical device provides a valuable solution for studying cell migration, facilitating a deeper understanding of cellular behaviors and the development of therapeutic strategies.

Introduction: Patient-derived organoids have emerged as a promising in vitro model for precision medicine, particularly in cancer, but also in noncancer-related diseases. However, the optimal culture medium for culturing patient-derived lung organoids has not yet been agreed upon. This study aimed to shed light on the optimal selection of a culture media for developing studies using patient-derived lung organoids.

Methods: Tumor and normal paired tissue from 71 resected non-small cell lung cancer patients were processed for organoid culture. Lung cancer organoids (LCOs) were derived from tumor tissue and normal lung organoids (LNOs) from nonneoplastic lung tissue. Three different culture media were compared: permissive culture medium (PCM), limited culture medium (LCM), and minimum basal medium (MBM). We assessed their effectiveness in establishing organoid cultures, promoting organoid growth and viability, and compared their differential phenotypic characteristics.

Results: While PCM was associated with the highest success rate and useful for long-term expansion, MBM was the best option to avoid normal organoid overgrowth in the organoid culture. The density, size, and viability of LNOs were reduced using LCM and severely affected with MBM. LNOs cultured in PCM tend to differentiate to bronchospheres, while alveolosphere differentiation can be observed in those cultured with LCM. The morphological phenotype of LCO was influenced by the culture media of election. Mesenchymal cell overgrowth was observed when LCM was used.

Conclusion: This work highlights the importance of considering the research objectives when selecting the most suitable culture medium for growing patient-derived lung organoids.

Introduction: Patient-derived organoids have emerged as a promising in vitro model for precision medicine, particularly in cancer, but also in noncancer-related diseases. However, the optimal culture medium for culturing patient-derived lung organoids has not yet been agreed upon. This study aimed to shed light on the optimal selection of a culture media for developing studies using patient-derived lung organoids.

Methods: Tumor and normal paired tissue from 71 resected non-small cell lung cancer patients were processed for organoid culture. Lung cancer organoids (LCOs) were derived from tumor tissue and normal lung organoids (LNOs) from nonneoplastic lung tissue. Three different culture media were compared: permissive culture medium (PCM), limited culture medium (LCM), and minimum basal medium (MBM). We assessed their effectiveness in establishing organoid cultures, promoting organoid growth and viability, and compared their differential phenotypic characteristics.

Results: While PCM was associated with the highest success rate and useful for long-term expansion, MBM was the best option to avoid n

Background: Paper-based microfluidics have gained significant attention as cost-effective and biocompatible platforms for various biological and medical applications. These devices facilitate the replication of complex tissue environments and offer a versatile alternative to traditional microfluidic systems.

Summary: This review highlights recent advances in paper-based microfluidics for tissue engineering and regenerative medicine. Key applications include 3D cell culture, bioanalysis assays, and high-throughput screening systems. Innovations in fabrication methods, such as wax printing and inkjet printing, have enhanced the functionality and scalability of these devices. Furthermore, the integration of biomaterials and surface modification techniques has improved their utility in replicating physiological conditions and studying cellular behaviors. Challenges such as mechanical robustness, imaging compatibility, and immune antigenicity are also addressed, alongside potential solutions and future directions.

Key messages: Paper-based microfluidic systems provide a transformative platform for tissue engineering and regenerative medicine, offering simplicity, affordability, and functional versatility. With ongoing innovations, these devices are poised to bridge the gap between laboratory research and clinical applications, supporting advancements in personalized medicine, regenerative therapies, and disease modeling.

Introduction: Mitochondria are cellular energy factories, but their function declines with age in many tissues as well as disease pathophysiology. Mitochondrial proteins have sugar modifications called glycans, which regulate their function and localization. There is a knowledge gap on the impact of mitochondrial protein glycosylation on mitochondrial function and mediating neuroinflammation. We hypothesize that stimuli-specific neuroinflammatory treatments in microglia induce pathological changes in mitochondrial protein glycosylation and compromise mitochondrial function.

Methods: The aim of this study was to establish a detailed microglial mitochondrial glycoprofile in different models of inflammation using lectins to identify the glycan-based markers of mitochondrial dysfunction. We use three different pathways of microglial activation: lipopolysaccharide, cytokines, and oxygen-glucose deprivation (OGD), revealing differences in mitochondrial glycosylation in different models of inflamed microglia. Mitochondrial lectin blots and lectin flow analysis were used to quantify the glycosylation changes due to different neuroinflammatory conditions. Seahorse Mito Stress assay was performed to assess mitochondrial function in each of these conditions.

Results: Lectin immunoblots of mitochondrial proteins and lectin flow studies with intact mitochondria were performed in three different neuroinflammation models using BV-2 microglial cells, revealing considerable stimuli-specific, differential mitochondrial glycosylation between these models and healthy controls. It was found that several glycans associated with mitochondria were differentially regulated during microglial activation. The observed changes in glycosylation trends were more drastic in OGD treatment as compared to other treatments, especially for complex and sialylated glycans.

Conclusion: This study represents the first functional investigation of mitochondrial glycosylation in microglial inflammation models towards identifying glycosylation-based therapeutic targets.

Introduction: Damage-associated molecular patterns (DAMPs) are molecules released in response to tissue or cellular damage to facilitate tissue regeneration. This inflammatory response can occur in sterile environments and is promoted by the release of damaged extracellular components such as the extracellular matrix (ECM). DAMPs have been implicated in various stages of wound healing but have yet to be explicitly utilized for regenerative medicine by leveraging selective modulation of the inflammatory response. With this in mind, we leverage inflammation to drive tissue regeneration by utilizing DAMPs collected from the native ECM, extracellular matrix motifs (mECM).

Methods: Here, mECMs were derived from UV-damaged rat tail collagen I. Fibroblast response to various concentrations and presentation of mECMs was investigated by evaluating changes in viability, proliferation, cell phenotype, and cytokine secretion.

Results: mECMs had reduced intensity in collagen I associated bands, indicating successful fragmentation to lower molecular weights. Soluble (mobile) mECMs induced changes in fibroblast phenotype as indicated by a decrease in proliferation, a decrease in nuclei area, and an increase in the percentage of elongated cells. In addition, mobile mECMs contributed to significant increases in cytokine secretion compared to insoluble (bound) mECMs. Across all experiments, bound mECMs exhibited effects on fibroblasts compared to the collagen control.

Conclusion: Fibroblasts in vitro recognize mECMs, with significant differences observed based on the presentation of these proteins. These data indicate that cryptic regions that are recognized by fibroblasts may be exposed in the mobile version of the mECMs, which lead to a myofibroblast-like phenotype in fibroblasts. This work highlights the potential of DAMPs to serve as immunomodulatory therapeutics for tissue regeneration.

Introduction: Fetal microchimerism could be involved in the regulation of breast cancer oncogenesis. CD34+ cells could be of a particular interest as up to 12% of the CD34+ population in maternal blood are of fetal origin. The aim of this research was to analyze the impact of umbilical cord blood (UCB) CD34+ on MCF-7 and MDA-MB-231 breast cancer cell lines, in order to uncover novel biological mechanisms and suggest novel treatment options for breast cancer.

Methods: UCB CD34+ cells were obtained from healthy women at full-term delivery. Direct cultures were grown with MCF-7 and MDA-MB-231 cells. Proliferation, migration, invasion, and transcriptomic analysis of breast cancer cell lines were compared between cultures exposed and nonexposed to UCB CD34+ cells. Interactions between UCB CD34+ and breast cancer cells were analyzed under fluorescent microscopy. Functional analyses were generated with QIAGEN's Ingenuity Pathway Analysis (IPA) and Gene Set Enrichment Analysis (GSEA).

Results: Direct contact between UCB CD34+ and breast cancer cell lines induced a reduction in the proliferative capacities of MCF-7 and MDA-MB-231 and diminished the migration abilities of MDA-MB-231 cells. In 3D coculture, UCB CD34+ cells were attracted by tumor spheroids and incorporated into tumor cells. These cell-to-cell interactions were responsible for transcriptome modifications coherent with observed functional modifications. Among the cytokines secreted by UCB CD34+, IFN-γ was identified as a potential upstream regulator responsible for the molecular modifications observed in transcriptomic analysis of MCF-7 breast cancer cells exposed to UCB CD34+ cells, as was IL-17A in MDA-MB-231 cells.

Conclusion: Direct cell-to-cell contact induced functional modifications in breast cancer cells. Interactions between UCB CD34+ and breast cancer cells could induce cell fusion and signal transmission via cytokines. Further analysis of direct cell-to-cell interactions should be performed at a molecular level to further understand the potential role of fetal CD34+ cells in breast cancer.

Introduction: Mesenchymal stem cell (MSC)-based therapies have emerged as a promising approach for treating articular cartilage injuries. However, enhancing the chondrogenic differentiation potential of MSCs remains a significant challenge. KDM6B, a histone demethylase that specifically removes H3K27me3 marks, is essential in controlling the maturation of chondrocytes. In this study, we examined how KDM6B influences chondrogenic differentiation in SCAPs and investigated the underlying mechanisms involved.

Methods: SCAPs were utilized. Alcian blue staining, pellet culture, and cell transplantation in rabbit knee cartilage defect models assessed MSC chondrogenic differentiation. Western blot, real-time RT-PCR, and microarray analysis examined the underlying molecular mechanisms.

Results: KDM6B promotes the expression of aggrecan, COL2A1, COL5, glycosaminoglycans, and collagen fibers, while also increasing the COL2/COL1 ratio in SCAPs. In vivo, SCAPs overexpressing KDM6B significantly enhanced the repair and regeneration of knee cartilage and subchondral bone, with higher levels of glycosaminoglycan and COL2 expression observed within the tissue. KDM6B promotes the chondrogenic differentiation potential of SCAPs by repressing HES1. In addition, knockdown of HES1 enhanced the chondrogenic differentiation of SCAPs.

Conclusions: KDM6B enhances the differentiation of SCAPs into chondrocytes and demonstrated its effectiveness in the repair and regeneration of cartilage tissue and subchondral bone in vivo experiments. These findings provide an important foundation for future research on the use of dental tissue-derived stem cells to treat cartilage injuries.

Introduction: The hedgehog signaling pathway plays a crucial role in inducing segment polarity through cell-cell interactions in various metazoans, including arthropods and annelids. However, its involvement in organogenesis and segmentation among lophotrochozoans remains inconsistent. This study aimed to explore the role of the hedgehog gene during gut development in the freshwater leech, Helobdella austinensis.

Methods: Developmental RT-PCR and in situ hybridization were performed to examine the expressions of hedgehog genes. In addition, embryos were treated with cyclopamine (a hedgehog signaling antagonist) and purmorphamine (a Smo agonist) to examine the potential interactions between Helobdella orthologs to hedgehog and two NKL genes: Hau-NK2 and Hau-NK4.

Results: We examined the expressions of four core pathway members - Hedgehog (Hh), Patched (Ptc), Smoothened (Smo), and the downstream transcription factor Gli - spatiotemporally during the embryonic stages of H. austinensis. All four genes were expressed in the developing gut and proboscis during organogenesis but not during the segmentation stage. Additionally, the treatment of embryos with cyclopamine and purmorphamine revealed that NK genes are regulated by hedgehog signaling. Furthermore, NK2 and NK4 were expressed in the developing gut rather than in a segmental stripe pattern.

Conclusion: This study confirms that the hedgehog signaling pathway is associated with gut development in the freshwater leech, H. austinensis. The expression patterns of hedgehog pathway genes and their interaction with NK genes suggest a role of hedgehog signaling in regulating gut development rather than segmentation in the freshwater leeches.

Introduction: To date, there have been no studies conducted on the development of interosseous muscles (IO) in the human hand. This study aimed to investigate the development of these muscles in order to clarify their terminal insertions and their relationship with the metacarpophalangeal joints.

Methods: Serial sections of 25 human specimens (9 embryos and 16 fetuses) between the 7th and 14th weeks of development, sourced from the Collection of the Department of Anatomy and Embryology at UCM Faculty of Medicine, were analyzed bilaterally using a conventional optical microscope.

Results: Our findings revealed that, during the 7th week of development, the metacarpophalangeal interzone mesenchyme extended into the extensor apparatus of the fingers. Furthermore, we observed that the joint capsule and the tendon of the IO derive from the articular interzone mesenchyme. By the end of the 7th week, corresponding to Carnegie stage 21, the myotendinous junction appeared, initiating cavitation of the metacarpophalangeal joint. During the fetal period, the terminal insertions of the IO were identified: both the dorsal interosseous (DI) and palmar interosseous (PI) muscles insert into the metacarpophalangeal joint capsule and establish a connection with the volar plate located at the base of the proximal phalanx and the extensor apparatus. Some muscle fibers also attach to the joint capsule at the level of the proximal synovial cul-de-sac. The functional implications of these findings are discussed within this work.

Conclusion: This study provides the first detailed description of the development of the interosseous muscles in the human hand.