Differentiation最新文献

{"title":"Evaluation of chondrogenesis and osteogenesis in human mesenchymal stem cells, chondrocytes, and chondroprogenitors using molecular markers, cellular markers and polarized microscopy","authors":"Archa Suresh ,&nbsp;Ayshath Ruksana C ,&nbsp;Ganesh Parasuraman ,&nbsp;Mohana Priya ,&nbsp;Grace Rebekah ,&nbsp;Christo Jeyaraj ,&nbsp;Elizabeth Vinod","doi":"10.1016/j.diff.2025.100898","DOIUrl":"10.1016/j.diff.2025.100898","url":null,"abstract":"<div><h3>Purpose</h3><div>Fibronectin adhesion assay progenitors (FAA-CPs) and migratory assay progenitors (MCPs), subsets of mesenchymal-like stromal cells (MSCs), exhibit superior in-vitro chondrogenic potential compared to bone marrow (BM)-MSCs and chondrocytes. To assess this potential, differentiation studies followed by confirmatory staining for collagen deposition are utilized. Polarized light microscopy (PLM), based on birefringence principles, is a valuable tool for visualizing organized collagen fibers. Its use as a predictive tool for assessing chondrogenesis and osteogenesis has not been reported.</div></div><div><h3>Methods</h3><div>This study involved FAA-CPs, MCPs, chondrocytes, and BM-MSCs derived from osteoarthritic knee joints (n = 3). After phenotypic characterization, the cells underwent chondrogenic and osteogenic differentiation, followed by Picrosirius red staining and PLM analysis, including immunohistochemical analysis for collagen types I, II, and X.</div></div><div><h3>Results</h3><div>Birefringence assessment revealed greater collagen fibril alignment and significant remodeling in the BM-MSC group, which exhibited an arcade-like pattern. The MCP group displayed well-organized collagen fibrils in pericellular zones and as a peripheral band, while chondrocytes and FAA-CPs exhibited lower intensity birefringence, indicating random alignment. Areas with higher collagen type II deposition corresponded to reduced collagen type I and the absence of collagen type X, highlighting the unique fibrillar network seen with PLM was indicative of collagen type II.</div></div><div><h3>Conclusion</h3><div>While its application for osteogenesis was limited, probably due to the non-fibrillar nature of collagen type X, its value for chondrogenesis is notable. Although not directly reflecting chondrogenesis, PLM can serve as a valuable tool for gaining insights into collagen remodeling, particularly concerning collagen type II during chondrogenic differentiation.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100898"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144722381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NONO regulates monocyte-macrophage lineage differentiation through a potential PI3K/AKT-dependent mechanism","authors":"Xiu-Rong Wei ,&nbsp;Dan Hu ,&nbsp;Zi-Jiang Yang ,&nbsp;Lv-Bin Yan ,&nbsp;Guang-Yu Xu ,&nbsp;Rui-Gang Zhang ,&nbsp;Xiu-Juan Zhang","doi":"10.1016/j.diff.2025.100901","DOIUrl":"10.1016/j.diff.2025.100901","url":null,"abstract":"<div><div>Non-POU domain containing octamer binding protein (NONO) is a multifunctional nuclear protein which plays important roles in regulating nuclear processes such as transcription and splicing. We aimed to delineate the effects and the underlying mechanisms of NONO on monocyte-macrophage lineage differentiation. By depolying a phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cell differentiation model and a macrophage colony-stimulating factor (M-CSF)-induced mouse bone marrow cell differentiation model, we examined the expression pattern and the effects of NONO during monocyte-macrophage lineage differentiation. The research revealed that the expression of NONO protein progressively decreased during the M-CSF-induced differentiation of mouse bone marrow cells into macrophages and the PMA-induced differentiation of THP-1 human monocytic leukemia cells into macrophages. The monocyte-macrophage lineage differentiation process was enhanced in <em>Nono</em> gene knockout (Nono K.O.) mouse bone marrow cells as well as <em>NONO</em> knockdown (NONO K.D.) THP-1 cells. The study also found that reduced NONO expression enhanced the AKT phosphorylation during macrophage lineage differentiation. At the same time, the PI3K inhibitor suppressed THP-1 cell differentiation into macrophages and attenuated the AKT phosphorylation activation by PMA and NONO knockdown during PMA-induced differentiation of THP-1 cells into macrophages. These results suggested an important role of NONO in regulating monocyte-macrophage lineage differentiation and this process was mediated, at least partially, through PI3K/AKT signaling pathway.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100901"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144895308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of Atg13-mediated autophagy enhances the anti-osteoclastogenic effect of sirolimus by counteracting its pro-autophagic activity","authors":"Tingwei Gao ,&nbsp;Jiankun Wang ,&nbsp;Jiajia Wang ,&nbsp;Xi Li ,&nbsp;Zhanhao Xiao","doi":"10.1016/j.diff.2025.100902","DOIUrl":"10.1016/j.diff.2025.100902","url":null,"abstract":"<div><div>Sirolimus can inhibit osteoclastogenesis. But sirolimus-activated autophagy is a favorable factor for osteoclastogenesis. This study aimed to explore the significance of autophagy in sirolimus-regulated osteoclastogenesis. Our results confirmed that sirolimus inhibited osteoclastic differentiation (including the number and size of osteoclasts as well as the expression of osteoclastic genes) and promotes osteoclast precursor (OCP) autophagy (including LC3 conversion and autophagosome/autolysosome formation). As expected, OCP autophagy (including LC3 conversion and LC3-puncta formation) promoted by sirolimus was reversed by autophagy inactivation with 3-MA or Atg13 silencing. Importantly, compared with single intervention of sirolimus, the combination of sirolimus and 3-MA or Atg13 silencing more effectively inhibited osteoclastic differentiation and OCP proliferation. <em>In vivo</em> experiments also demonstrated that the combination of sirolimus and Atg13-silencing adeno-associated viruses (AAVs) was more effective than sirolimus alone in improving decreased bone density and damaged bone microstructure (including Micro-CT imaging results, bone tissue parameters and trabecular area), and attenuating osteoclastic activity (including the abundance of osteoclasts in trabecular bones and the production of osteoclastic markers in serum) in ovariectomized (OVX) mice. In conclusion, repressing Atg13-related autophagy can effectively enhance the function of sirolimus in inhibiting osteoclastogenesis by counteracting its pro-autophagic activity. Therefore, the combination of sirolimus and Atg13-targeting therapy is expected to enhance the efficacy of sirolimus in ameliorating osteoclastic osteoporosis.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100902"},"PeriodicalIF":2.6,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144903192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enteric neural crest development in Astyanax mexicanus surface fish and cavefish","authors":"Pavani Ponnimbaduge Perera ,&nbsp;Kaitlyn Webster ,&nbsp;Misty R. Riddle","doi":"10.1016/j.diff.2025.100881","DOIUrl":"10.1016/j.diff.2025.100881","url":null,"abstract":"<div><div>The enteric nervous system (ENS) regulates gastrointestinal (GI) functions such as secretion, blood flow, and motility, yet how its structure and function evolve with dietary adaptations remains unclear. <em>Astyanax mexicanus</em>, a teleost fish with surface and cave morphotypes, offers a model to explore these changes; cavefish exhibit altered GI motility and transit that may help them adapt to their unique diet. We compared early ENS development in surface fish and cavefish, tracking enteric neural crest cell (ENCC) migration and differentiation using <em>phox2bb</em> and HuC/D markers. We found that ENCCs reach the gut by 36 hours post-fertilization (hpf) in both morphotypes but migrate and differentiate more rapidly along the gut in cavefish. To explore the genetic basis of this difference, we used available genomic datasets to compare the predicted peptide sequences of genes important for ENS development in other species and identified mutations that could impact protein function, for example in the endothelin signaling genes important for ENCC migration and differentiation. We specifically examined the expression of <em>endothelin-3 (edn3)</em> and <em>endothelin receptor-b a (ednrba)</em> during ENCC migration and found that the localization of <em>edn3</em>, but not <em>ednrba</em>, is consistent with a potential role in ENS development. Overall, our findings establish <em>A. mexicanus</em> as a model for studying evolution of ENS development.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100881"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144365759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Developmental biology in Chile: an open window to Latin America and the world","authors":"Miguel L. Concha ,&nbsp;Marcia Gaete ,&nbsp;Sylvain Marcellini ,&nbsp;Verónica Palma ,&nbsp;Marcela Torrejón ,&nbsp;Constanza Vásquez-Doorman","doi":"10.1016/j.diff.2025.100882","DOIUrl":"10.1016/j.diff.2025.100882","url":null,"abstract":"<div><div>The field of developmental biology has a relatively short history in Chile. It began with the pioneering work of Luis Izquierdo in the 1950s, when he challenged the prevailing views on embryology and comparative anatomy. Since then, the developmental biology community has grown considerably, broadening its vision and perspectives, as well as its international presence. Within the Latin American context, Chile has played a leading role in promoting the discipline and training a new generation of developmental biologists. In 2024, the country hosted several impactful regional events including a hands-on course, an international conference, a global lecture series, and the inaugural congress of the Chilean Society for Developmental Biology (SBDCh). The pivotal role played by the European Molecular Biology Organisation (EMBO), the United Nations University Programme for Biotechnology in Latin America and the Caribbean (UNU-BIOLAC), and the International Society of Differentiation (ISD) in supporting Chile's leadership in the field is also worth highlighting. These organisations enabled the participation of young Latin American researchers by providing travel grants and support. This is particularly relevant in today's changing world, where global integration and collaboration are fundamental to the effective promotion of developmental biology, especially among younger generations.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100882"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144481047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of Pumilio gene expression during early neural crest development","authors":"Mariann Guzman-Espinoza ,&nbsp;Helen M. Vander Wende ,&nbsp;Jessica L. Pacheco ,&nbsp;Alejandra Olano Roldán ,&nbsp;Erica J. Hutchins","doi":"10.1016/j.diff.2025.100883","DOIUrl":"10.1016/j.diff.2025.100883","url":null,"abstract":"<div><div>Neural crest cells are multipotent cells present in vertebrate embryos that give rise to a wide array of cell types and tissues. A growing number of studies have identified post-transcriptional regulatory events that are essential for multiple stages of neural crest development, though a thorough characterization of the post-transcriptional regulators controlling these events is currently lacking. From single cell RNA-sequencing data, we identified members of the Pumilio family of RNA-binding proteins, PUM1 and PUM2, as candidate post-transcriptional regulators of neural crest development. Using hybridization chain reaction (HCR) in avian embryos (<em>Gallus gallus</em>), we characterized the spatiotemporal expression of <em>Pumilio</em> family mRNAs during early stages of cranial neural crest development. We show that <em>Pum1</em> and <em>Pum2</em>, though expressed throughout the three germ layers, were enriched in ectodermally-derived tissues, and following neurulation, <em>Pum1</em> and <em>Pum2</em> show distinct expression patterns. We observed that <em>Pum1</em> displayed a more uniform expression throughout the neural tube and neural crest during neural crest specification and the epithelial-mesenchymal transition (EMT). In contrast, <em>Pum2</em> was enriched in neural crest cells poised to undergo EMT. We thus hypothesize that PUM1 and PUM2, often speculated to be functionally redundant, may play distinct roles at key steps of neural crest development.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100883"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144489521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Shared and individual expression patterns of pluripotency genes in the developing chick embryo during neurulation and beyond","authors":"Jenny Hsin ,&nbsp;Rita M. Yazejian ,&nbsp;Ceren Pajanoja ,&nbsp;Laura Kerosuo","doi":"10.1016/j.diff.2025.100866","DOIUrl":"10.1016/j.diff.2025.100866","url":null,"abstract":"<div><div>The neural crest (NC) is a transient population of pluripotent-like, pleistopotent stem cells that emerges early in vertebrate development. These cells play a pivotal role in generating a diverse array of tissues, including the craniofacial bone and cartilage, the entire peripheral nervous system, melanocytes of the skin, certain cardiac structures, and chromaffin cells of the adrenal medulla, among others. The stem cell potential of neural crest cells (NCCs) has long intrigued developmental biologists, as the NC originates post-gastrulation in the ectoderm, yet NCCs also give rise to derivatives typically associated with mesodermal or endodermal origins. Recent work has shown that NCCs co-express factors known from the core pluripotency complex from the pre-gastrulation stages in the epiblast, which enables their exceptionally high stem cell potential. However, detailed spatiotemporal data on pluripotency factor expression in vertebrate embryos remain limited, and the distinction between the function of co-expression of pluripotency genes versus their individual expression in the developing embryo is not clear. In this study, to elucidate the NCC formation process across axial levels as well as the putative different roles of these stem cell genes during early embryogenesis, we used multi-channel fluorescent in situ hybridization to comprehensively examine the anterior-to-posterior expression of pluripotency factors PouV (Oct4), Nanog, Klf4 and Lin28A in chick embryos across key developmental stages, from Hamburger and Hamilton (HH) stage 5 to stage 14. From head to trunk, we find that while the early ectoderm, including the future epidermis and central nervous system (CNS) domains, in the neural fold stages broadly co-express these genes, their expression profiles differ significantly after neurulation. Nanog expression remains in the hindbrain and vagal migratory NCCs. Klf4 strongly marks the developing floor plate, and Klf4, PouV and Lin28A are expressed also in the neural tube that forms the CNS as well as in the developing somites, implying additional roles for these factors during embryogenesis.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100866"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144177534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Origin stories of neural crest roles in craniofacial development: A tale of the meninges","authors":"Diana Rigueur ,&nbsp;Julia C. Boughner","doi":"10.1016/j.diff.2025.100880","DOIUrl":"10.1016/j.diff.2025.100880","url":null,"abstract":"<div><div>As developmental biologists, we tell origin stories to explain differences in vertebrate craniofacial form and complexity. Some of these differences are linked to diseases, notably of patients born with congenital malformations, many of them syndromic. These defective processes impede proper neural crest cell migration, differentiation, and patterning of bones, cartilage, joints, muscle and other connective tissues, including of the head. For example, a rare form of holoprosencephaly called cyclopia is a particular quandary in its diverse presentation. This syndromic congenital disease includes lost brain frontal lobe septation and development of a single eye. Subsequent craniofacial midline defects include frontal suture synostosis, absent nose and medial maxilla, and a restructuring of facial bones to accommodate one eye. The meningeal tissues in contact with both the underlying brain and the overlying calvarial bone are derived at least in part from neural crest cells. This Perspectives piece re-examines communication between the neural crest cell-brain axis, explores the direct involvement of the meninges in this communication process, and proposes an expanded origin story about the etiology of craniofacial patterning and disease.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100880"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144306320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Myogenic and adipogenic potential of porcine muscle satellite cells isolated by flow cytometry","authors":"A. Vincent ,&nbsp;K. Stange ,&nbsp;I. Louveau ,&nbsp;M. Röntgen ,&nbsp;F. Dessauge","doi":"10.1016/j.diff.2025.100871","DOIUrl":"10.1016/j.diff.2025.100871","url":null,"abstract":"<div><div>Skeletal muscle mesenchymal stromal cells (MSCs) are resident cells in the stromal, interstitial or perivascular areas, with satellite cells (SCs) acting as essential precursors for muscle growth and regeneration. This study firstly aimed to phenotype freshly isolated SCs using cell surface markers and gene expression and to assess their ability to differentiate in myogenic or adipogenic conditions. Then, refined SCs populations sorted according to the myogenic CD56 cell surface marker were characterized. SCs were isolated from the l<em>ongissimus</em> muscle of 5-7-day-old piglets and analyzed using flow cytometry. The hematopoietic CD45 + cells represented 20 % of the total isolated cell population. The myogenic CD29 and CD56 positive cell populations were the most abundant (80 % and 50 % respectively). Mesenchymal CD90 positive cells were also highly present (34 %) while the proportions of fibro-adipogenic CD140a and CD34-positive cells were low (&lt;3 %). In this study, we showed that total isolated muscle-derived SCs were able to differentiate into myotubes in both myogenic and adipogenic media. Transcriptional profiles were similar, except for PPARγ, PGC1α, and Myosin Heavy Chain 2B witch present a higher induction during differentiation in the adipogenic medium. Interestingly, non-hematopoietic CD45⁻sorted cells further separated in CD45-/CD56+ and also CD45-/CD56-sub-populations mostly formed myotubes under both conditions, with CD56⁻cells showing potential and gene expression profile of myoblasts. Mature adipocytes were found in the CD45-/CD56+ group after differentiation in adipogenic medium. This study increases knowledge on myogenic cell surface marker and underscores the complexity and heterogeneity of muscle SCs.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100871"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144169608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “The mechanism of EGF in promoting skeletal muscle post-injury regeneration” [Differentiation 143 (2025) 100862]","authors":"Huaixin Teng ,&nbsp;Yongze Liu ,&nbsp;Ruotong Hao ,&nbsp;Lu Zhang ,&nbsp;Xiaoyu Zhang ,&nbsp;Shufeng Li ,&nbsp;Shuang Li ,&nbsp;Huili Tong","doi":"10.1016/j.diff.2025.100885","DOIUrl":"10.1016/j.diff.2025.100885","url":null,"abstract":"","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100885"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144522185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}