Pub Date : 2025-06-23DOI: 10.1016/j.diff.2025.100883
Mariann Guzman-Espinoza , Helen M. Vander Wende , Jessica L. Pacheco , Alejandra Olano Roldán , Erica J. Hutchins
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 (Gallus gallus), we characterized the spatiotemporal expression of Pumilio family mRNAs during early stages of cranial neural crest development. We show that Pum1 and Pum2, though expressed throughout the three germ layers, were enriched in ectodermally-derived tissues, and following neurulation, Pum1 and Pum2 show distinct expression patterns. We observed that Pum1 displayed a more uniform expression throughout the neural tube and neural crest during neural crest specification and the epithelial-mesenchymal transition (EMT). In contrast, Pum2 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.
{"title":"Characterization of Pumilio gene expression during early neural crest development","authors":"Mariann Guzman-Espinoza , Helen M. Vander Wende , Jessica L. Pacheco , Alejandra Olano Roldán , 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":"2025-06-23","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}
Pub Date : 2025-06-16DOI: 10.1016/j.diff.2025.100882
Miguel L. Concha , Marcia Gaete , Sylvain Marcellini , Verónica Palma , Marcela Torrejón , Constanza Vásquez-Doorman
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.
{"title":"Developmental biology in Chile: an open window to Latin America and the world","authors":"Miguel L. Concha , Marcia Gaete , Sylvain Marcellini , Verónica Palma , Marcela Torrejón , 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":"2025-06-16","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}
Pub Date : 2025-06-13DOI: 10.1016/j.diff.2025.100881
Pavani Ponnimbaduge Perera , Kaitlyn Webster , Misty R. Riddle
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. Astyanax mexicanus, 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 phox2bb 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 endothelin-3 (edn3) and endothelin receptor-b a (ednrba) during ENCC migration and found that the localization of edn3, but not ednrba, is consistent with a potential role in ENS development. Overall, our findings establish A. mexicanus as a model for studying evolution of ENS development.
肠神经系统（ENS）调节胃肠道（GI）功能，如分泌、血流和运动，但其结构和功能如何随着饮食适应而进化尚不清楚。Astyanax mexicanus，一种具有表面和洞穴形态的硬骨鱼，为探索这些变化提供了一个模型；洞穴鱼表现出改变的胃肠道运动和运输，这可能有助于它们适应独特的饮食。我们比较了表层鱼和洞穴鱼的早期ENS发育，利用phox2bb和HuC/D标记跟踪了肠神经嵴细胞（enteric neural crest cell， ENCC）的迁移和分化。我们发现，两种形态的ENCCs在受精后36小时到达肠道，但在洞穴鱼中，ENCCs沿着肠道迁移和分化的速度更快。为了探索这种差异的遗传基础，我们使用现有的基因组数据集来比较其他物种ENS发育重要基因的预测肽序列，并确定可能影响蛋白质功能的突变，例如对ENCC迁移和分化重要的内皮素信号基因。我们专门检查了内皮素-3 （edn3）和内皮素受体-b - a （ednrba）在ENCC迁移过程中的表达，发现edn3的定位，而不是ednrba，与ENS发展的潜在作用一致。总的来说，我们的研究结果建立了墨西哥拟南猿作为研究ENS发展进化的模型。
{"title":"Enteric neural crest development in Astyanax mexicanus surface fish and cavefish","authors":"Pavani Ponnimbaduge Perera , Kaitlyn Webster , 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":"2025-06-13","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}
Pub Date : 2025-06-05DOI: 10.1016/j.diff.2025.100880
Diana Rigueur , Julia C. Boughner
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.
{"title":"Origin stories of neural crest roles in craniofacial development: A tale of the meninges","authors":"Diana Rigueur , 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":"2025-06-05","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}
Pub Date : 2025-05-29DOI: 10.1016/j.diff.2025.100872
Jorge Alejandro Barbosa-Nuñez , José Nabor Haro-González , Eristeo García-Márquez , Hugo Espinosa-Andrews , Eduardo Padilla-Camberos , Sara Elisa Herrera-Rodríguez
Bones are living tissues that are periodically renewed through the bone remodeling process. It starts when monocyte-macrophage-type cells begin to proliferate and recruit into the bone tissue due to the action of various proteins. The proteins are generated by the body in response to a lack of calcium in the blood or a damaged bone tissue. Then, other chemical signals promote the differentiation of monocyte-macrophage-type cells into osteoclasts. Finally, osteoclasts carry out a process known as bone resorption in which they degrade bone tissue. After, bone resorption a process known as osteogenesis takes place. In osteogenesis, pluripotent mesenchymal stem cells proliferate and recruit in the bone damaged by bone resorption in response to the action of different compounds. Then, pluripotent mesenchymal stem cells differentiate into osteoblasts due to the effect of specific proteins. Finally, osteoblasts generate new bone tissue, completing the bone remodeling process. This review aims to summarize and provide recent findings of the proteins and chemical signals involved in osteoclast and osteoblast proliferation, recruitment and maturation during the bone resorption and osteogenesis processes. The information found about these molecules could help to better understand all the complex mechanisms that directly or indirectly influence the bone remodeling process. In this sense, the mechanisms of different bone disorders and diseases could be elucidated in a better way, leading to the generation of efficient and specific treatments for each of them in the future.
{"title":"Proteins and peptides responsible for bone remodeling","authors":"Jorge Alejandro Barbosa-Nuñez , José Nabor Haro-González , Eristeo García-Márquez , Hugo Espinosa-Andrews , Eduardo Padilla-Camberos , Sara Elisa Herrera-Rodríguez","doi":"10.1016/j.diff.2025.100872","DOIUrl":"10.1016/j.diff.2025.100872","url":null,"abstract":"<div><div>Bones are living tissues that are periodically renewed through the bone remodeling process. It starts when monocyte-macrophage-type cells begin to proliferate and recruit into the bone tissue due to the action of various proteins. The proteins are generated by the body in response to a lack of calcium in the blood or a damaged bone tissue. Then, other chemical signals promote the differentiation of monocyte-macrophage-type cells into osteoclasts. Finally, osteoclasts carry out a process known as bone resorption in which they degrade bone tissue. After, bone resorption a process known as osteogenesis takes place. In osteogenesis, pluripotent mesenchymal stem cells proliferate and recruit in the bone damaged by bone resorption in response to the action of different compounds. Then, pluripotent mesenchymal stem cells differentiate into osteoblasts due to the effect of specific proteins. Finally, osteoblasts generate new bone tissue, completing the bone remodeling process. This review aims to summarize and provide recent findings of the proteins and chemical signals involved in osteoclast and osteoblast proliferation, recruitment and maturation during the bone resorption and osteogenesis processes. The information found about these molecules could help to better understand all the complex mechanisms that directly or indirectly influence the bone remodeling process. In this sense, the mechanisms of different bone disorders and diseases could be elucidated in a better way, leading to the generation of efficient and specific treatments for each of them in the future.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"144 ","pages":"Article 100872"},"PeriodicalIF":2.2,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144189633","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}
Pub Date : 2025-05-27DOI: 10.1016/j.diff.2025.100871
A. Vincent , K. Stange , I. Louveau , M. Röntgen , F. Dessauge
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 longissimus 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 (<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.
{"title":"Myogenic and adipogenic potential of porcine muscle satellite cells isolated by flow cytometry","authors":"A. Vincent , K. Stange , I. Louveau , M. Röntgen , 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 (<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<sup>−</sup>sorted cells further separated in CD45-/CD56+ and also CD45-/CD56-sub-populations mostly formed myotubes under both conditions, with CD56<sup>−</sup>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":"2025-05-27","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}
Pub Date : 2025-05-15DOI: 10.1016/j.diff.2025.100866
Jenny Hsin , Rita M. Yazejian , Ceren Pajanoja , Laura Kerosuo
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.
{"title":"Shared and individual expression patterns of pluripotency genes in the developing chick embryo during neurulation and beyond","authors":"Jenny Hsin , Rita M. Yazejian , Ceren Pajanoja , 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":"2025-05-15","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}
Pub Date : 2025-05-01DOI: 10.1016/j.diff.2025.100870
Yourka D. Tchoukalova , Manisha K. Shah , Cheryl E. Myers , Nan Zhang , David G. Lott
This study investigates the biological factors influencing the epithelial differentiation of adipose-derived stem cells (ASC) to develop an engineered upper airway construct. One fraction of ASC was seeded onto a fibrin sealant (Tisseel) matrix encompassing an additional equal fraction of ASC that has been integrated into a porous polyethylene scaffold (Medpor®). Constructs with ASC seeded at total densities of 5 × 105, 1 × 106, 2.5 × 106, and 5 × 106 cells cm-2 were cultured under submerged conditions for 11 days to achieve partial epithelial differentiation (PD). To simulate post-transplantation exposure to air and interaction with host epithelial cells, PD constructs with ASC at 5 × 106 cells cm-2 were transitioned to air-liquid interface (ALI) conditions for additional 10 days (PD/ALI-21d) or 21 days (PD/ALI-32d). The latter cultures were either maintained alone or co-cultured with bronchial epithelial cells (PD/ALI-32d + BEAS). Gene expressions of mesenchymal and epithelial basal, secretory, and ciliated cell markers were assessed and validated via immunohistochemistry.
ASC seeded at 5 × 106 cells cm-2 achieved the highest partial epithelial differentiation, supporting the use of this density for further experiments. In PD/ALI-21d, basal and secretory epithelial marker gene expression significantly increased, while ciliated cell markers remained unchanged. In PD/ALI-32d, expression of basal and goblet cell markers and several mesenchymal stem cell markers decreased, but co-culturing with BEAS maintained the levels of their expression. These results indicate that long-term ALI cultures cannot sustain terminal differentiation of ASC into secretory phenotypes without co-culture with primary epithelial cells.
In conclusion, partially differentiated ASC on constructs maintain a stem cell phenotype and may promote differentiation into basal/secretory phenotypes, but not ciliated cells. Enhancing ciliogenesis and ensuring ASC commitment to the epithelial lineage, require modifications to the study design.
{"title":"Optimization of seeding cell density for differentiation of adipose-derived stem cells into epithelial-like cells on bioengineered composite scaffolds","authors":"Yourka D. Tchoukalova , Manisha K. Shah , Cheryl E. Myers , Nan Zhang , David G. Lott","doi":"10.1016/j.diff.2025.100870","DOIUrl":"10.1016/j.diff.2025.100870","url":null,"abstract":"<div><div>This study investigates the biological factors influencing the epithelial differentiation of adipose-derived stem cells (ASC) to develop an engineered upper airway construct. One fraction of ASC was seeded onto a fibrin sealant (Tisseel) matrix encompassing an additional equal fraction of ASC that has been integrated into a porous polyethylene scaffold (Medpor®). Constructs with ASC seeded at total densities of 5 × 10<sup>5</sup>, 1 × 10<sup>6</sup>, 2.5 × 10<sup>6</sup>, and 5 × 10<sup>6</sup> cells cm-2 were cultured under submerged conditions for 11 days to achieve partial epithelial differentiation (PD). To simulate post-transplantation exposure to air and interaction with host epithelial cells, PD constructs with ASC at 5 × 10<sup>6</sup> cells cm-2 were transitioned to air-liquid interface (ALI) conditions for additional 10 days (PD/ALI-21d) or 21 days (PD/ALI-32d). The latter cultures were either maintained alone or co-cultured with bronchial epithelial cells (PD/ALI-32d + BEAS). Gene expressions of mesenchymal and epithelial basal, secretory, and ciliated cell markers were assessed and validated via immunohistochemistry.</div><div>ASC seeded at 5 × 10<sup>6</sup> cells cm-2 achieved the highest partial epithelial differentiation, supporting the use of this density for further experiments. In PD/ALI-21d, basal and secretory epithelial marker gene expression significantly increased, while ciliated cell markers remained unchanged. In PD/ALI-32d, expression of basal and goblet cell markers and several mesenchymal stem cell markers decreased, but co-culturing with BEAS maintained the levels of their expression. These results indicate that long-term ALI cultures cannot sustain terminal differentiation of ASC into secretory phenotypes without co-culture with primary epithelial cells.</div><div>In conclusion, partially differentiated ASC on constructs maintain a stem cell phenotype and may promote differentiation into basal/secretory phenotypes, but not ciliated cells. Enhancing ciliogenesis and ensuring ASC commitment to the epithelial lineage, require modifications to the study design.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"143 ","pages":"Article 100870"},"PeriodicalIF":2.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144123975","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}
Pub Date : 2025-05-01DOI: 10.1016/j.diff.2025.100865
Matthew J. Anderson, Mark Lewandoski
{"title":"Response to Wu et al","authors":"Matthew J. Anderson, Mark Lewandoski","doi":"10.1016/j.diff.2025.100865","DOIUrl":"10.1016/j.diff.2025.100865","url":null,"abstract":"","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"143 ","pages":"Article 100865"},"PeriodicalIF":2.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143928965","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}
Pub Date : 2025-05-01DOI: 10.1016/j.diff.2025.100867
Catharina Marques Sacramento, Márcio Zaffalon Casati, Enilson Antonio Sallum, Renato Corrêa Viana Casarin, Karina Gonzales Silvério
Background
The relationship between gastrointestinal hormones and bone metabolism has gained significant attention, but the specific role of cholecystokinin (CCK) in bone homeostasis remains largely unexplored. This study aimed to evaluate the role of the CCK pathway in osteogenic differentiation by blocking its mechanisms in human bone marrow stem cells (hBMSCs).
Methods
hBMSCs were exposed to Lorglumide, a CCK signaling pathway inhibitor, under osteogenic conditions. Cell viability, osteogenic differentiation, RT-qPCR analysis of CCK, FOS, OCN, and RUNX2, IP3 receptor phosphorylation, alkaline phosphatase (ALP) activity, and calcium concentration (Ca2) were assessed to elucidate Lorglumide's effects on osteogenesis and related mechanisms.
Results
Lorglumide reduced hBMSC viability at concentrations ≥30 μM over 14 days. Mineralization assays revealed dose-dependent inhibition, with 20 μM maintaining mineralization comparable to controls. RT-qPCR showed that Lorglumide suppressed CCK expression and altered osteogenic gene expression (FOS, RUNX2, OCN). Lorglumide decreased Ca2 concentration compared to osteogenic medium (OM) and reduced ALP activity, indicating its inhibitory effect on key osteogenic mechanisms.
Conclusion
Lorglumide inhibits hBMSC osteoblastic differentiation, suggesting a possible role for the CCK signaling pathway in bone metabolism. These findings emphasize the involvement of gastrointestinal hormones in bone homeostasis, suggesting new therapeutic opportunities targeting hormonal regulation to promote bone health. Further studies are needed to explore the underlying mechanisms and potential clinical applications of modulating CCK pathways in bone-related disorders.
{"title":"Cholecystokinin-antagonist lorglumide inhibits osteogenic differentiation in human bone marrow stem cells","authors":"Catharina Marques Sacramento, Márcio Zaffalon Casati, Enilson Antonio Sallum, Renato Corrêa Viana Casarin, Karina Gonzales Silvério","doi":"10.1016/j.diff.2025.100867","DOIUrl":"10.1016/j.diff.2025.100867","url":null,"abstract":"<div><h3>Background</h3><div>The relationship between gastrointestinal hormones and bone metabolism has gained significant attention, but the specific role of cholecystokinin (CCK) in bone homeostasis remains largely unexplored. This study aimed to evaluate the role of the CCK pathway in osteogenic differentiation by blocking its mechanisms in human bone marrow stem cells (hBMSCs).</div></div><div><h3>Methods</h3><div>hBMSCs were exposed to Lorglumide, a CCK signaling pathway inhibitor, under osteogenic conditions. Cell viability, osteogenic differentiation, RT-qPCR analysis of CCK, FOS, OCN, and RUNX2, IP3 receptor phosphorylation, alkaline phosphatase (ALP) activity, and calcium concentration (Ca<sup>2</sup>) were assessed to elucidate Lorglumide's effects on osteogenesis and related mechanisms.</div></div><div><h3>Results</h3><div>Lorglumide reduced hBMSC viability at concentrations ≥30 μM over 14 days. Mineralization assays revealed dose-dependent inhibition, with 20 μM maintaining mineralization comparable to controls. RT-qPCR showed that Lorglumide suppressed CCK expression and altered osteogenic gene expression (FOS, RUNX2, OCN). Lorglumide decreased Ca<sup>2</sup> concentration compared to osteogenic medium (OM) and reduced ALP activity, indicating its inhibitory effect on key osteogenic mechanisms.</div></div><div><h3>Conclusion</h3><div>Lorglumide inhibits hBMSC osteoblastic differentiation, suggesting a possible role for the CCK signaling pathway in bone metabolism. These findings emphasize the involvement of gastrointestinal hormones in bone homeostasis, suggesting new therapeutic opportunities targeting hormonal regulation to promote bone health. Further studies are needed to explore the underlying mechanisms and potential clinical applications of modulating CCK pathways in bone-related disorders.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"143 ","pages":"Article 100867"},"PeriodicalIF":2.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144068118","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}
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