Acta histochemica最新文献

{"title":"Kisspeptin (Kp-10) inhibits in vitro osteogenic differentiation of multipotent mesenchymal stromal cells extracted from the bone marrow of adult rats","authors":"Laís Bitencourt Guimarães ,&nbsp;Daniel Portela Dias Machado ,&nbsp;Beatriz Ferreira Carvalho Versiani Caldeira ,&nbsp;Larissa Tiemi Matuzake Vieira ,&nbsp;Gabriela Alves Santos ,&nbsp;Fabiana Rocha Araújo ,&nbsp;Leonardo Teotônio Machado ,&nbsp;Dawidson Assis Gomes ,&nbsp;Natália de Melo Ocarino ,&nbsp;Rogéria Serakides ,&nbsp;Amanda Maria Sena Reis","doi":"10.1016/j.acthis.2023.152112","DOIUrl":"10.1016/j.acthis.2023.152112","url":null,"abstract":"<div><p>Kisspeptin (Kp-10) is a neuropeptide that binds to GPR54 receptors, exerting several functions mainly in the nervous and reproductive systems of the body. However, its effects and mechanisms of action on the skeletal system remain poorly understood. This study evaluated the effects of different concentrations of Kp-10 on <em>in vitr</em>o osteogenic differentiation of multipotent mesenchymal stromal cells (MSCs) extracted from the bone marrow (BM) of adult Wistar rats. Two-month-old female rats were euthanized to extract BM from long bones to obtain MSCs. Four experimental groups were established <em>in vitro</em>: a control and Kp-10 at concentrations of 0.01, 0.05 and, 0.1 µg/mL. After induction of osteogenic differentiation, cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)− 2,5-diphenyl tetrazolium bromide (MTT) assay, alkaline phosphatase activity, collagen synthesis, percentage of area covered by MSCs/field and mineralized nodules/field, and immunocytochemistry of the GPR54 receptor tests. Furthermore, evaluation of gene transcripts for type I collagen, <em>Runx-2</em>, Bmp-2<em>,</em> bone sialoprotein, osteocalcin and osteopontin was performed using real-time RT-qPCR. It was observed that MSCs expressed GPR54 receptor to which Kp-10 binds during osteogenic differentiation, promoting a negative effect on osteogenic differentiation. This effect was observed at all the Kp-10 concentrations in a concentration-dependent manner, characterized by a decrease in the activity of alkaline phosphatase, collagen synthesis, mineralized nodules, and decreased expression of gene transcripts for type I collagen, osteocalcin, osteopontin, and <em>Runx-2</em>. Thus, Kp-10 inhibits <em>in vitro</em> osteogenic differentiation of MSCs extracted from the BM of adult Wistar rats.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152112"},"PeriodicalIF":2.5,"publicationDate":"2023-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72207931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In vitro validation: GLY alleviates UV-induced corneal epithelial damage through the HMGB1-TLR/MyD88-NF-κB signaling pathway","authors":"XinYi Chen ,&nbsp;XiaoXiao Zheng ,&nbsp;Ting Shen ,&nbsp;Ting He ,&nbsp;YangQi Zhao ,&nbsp;Yi Dong","doi":"10.1016/j.acthis.2023.152111","DOIUrl":"10.1016/j.acthis.2023.152111","url":null,"abstract":"<div><p>UV-induced corneal damage is a common ocular surface injury that usually leads to corneal lesions causing persistent inflammation. High mobility group box 1 (HMGB1) is identified as an inflammatory alarm in various tissue injuries. Here, this study first evaluates the repair effect of the HMGB1-selective inhibitor GLY in UV-induced corneal damage; Secondly, the inhibitory effect of GLY on UV-induced corneal damage induced inflammation and the potential therapeutic mechanism of GLY were studied. GLY effectively attenuates the expression of UV-induced inflammatory factors and HMGB1, TLR/MyD88, NF-κB signaling pathway genes at the mRNA and protein levels. In addition, RT-PCR and Western Blot experiments after knocking down HMGB1 and TLR2/9 genes showed that GLY alleviated corneal inflammation by inhibiting the HMGB1-TLR/MyD88 signaling pathway. The results of this study show that targeting HMGB1-NF-κB by GLY can alleviate the inflammatory response induced by UV induction.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152111"},"PeriodicalIF":2.5,"publicationDate":"2023-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71520158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect on m6A methylation writer complex by the reduced MATR3 in pterygium","authors":"Qianqian Guo,&nbsp;Shichao Han","doi":"10.1016/j.acthis.2023.152101","DOIUrl":"https://doi.org/10.1016/j.acthis.2023.152101","url":null,"abstract":"<div><p><span><span>Pterygium is a common eye surface disease with high recurrence and unclear pathogenesis. In current study, </span>RNA sequencing was conducted in 6 pairs of human pterygium and conjunctival tissues, and </span><em>Matr3</em><span><span><span> as a novel candidate gene was significantly reduced in pterygium compared to control tissues. Moreover, immunoprecipitation was performed to pull down MATR3, and WTAP specially interacting with MATR3 in control but not pterygium was identified by mass spectrum. Immunoprecipitation was performed to validate the interaction between MATR3 and WTAP/METTL3/METTL14 complex. (Methylated) </span>RNA<span> immunoprecipitation was performed to further reveal that the binding affinity of WTAP and MATR3 was lost at 3′ UTR of RNA molecules of down-regulated genes in pterygium. Overall, we figured out the loss of intercrossing between MATR3 and N6-methyladenosine </span></span>methyltransferase complex, as well as indicated the potential impact on transcription of target genes in pterygium.</span></p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152101"},"PeriodicalIF":2.5,"publicationDate":"2023-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91991208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MiR-223–3p overexpressed adipose mesenchymal stem cell-derived exosomes promote wound healing via targeting MAPK10","authors":"Xiaojiao Liu ,&nbsp;Shunqiao Jin ,&nbsp;Jiao Liu ,&nbsp;Xuezhu Xu","doi":"10.1016/j.acthis.2023.152102","DOIUrl":"10.1016/j.acthis.2023.152102","url":null,"abstract":"<div><h3>Background</h3><p><span>Adipose mesenchymal stem cell (AMSC)-derived exosomes are promising novel factors for wound repair and regeneration. This study aimed to explore the potential roles and underlying mechanisms of specific </span>miRNA in wound healing using AMSC-derived exosomes as carriers.</p></div><div><h3>Methods</h3><p>The expression profiles of GSE197840 were downloaded to screen for differentially expressed miRNAs (DEmiRNAs), and the corresponding genes of the identified miRNAs were predicted. Next, miRNA-mRNA co-expression networks were constructed and the genes in these networks were subjected to functional analysis. miR-223–3p overexpressed AMSCs were then established to isolate exosomes, and the effects of AMSC-derived exosomes carrying miR-223–3p on wound healing and the related potential mechanisms were further investigated in vivo.</p></div><div><h3>Results</h3><p><span><span><span>35 DEmiRNAs were identified and a co-expression network containing 22 miRNAs and 91 target genes was constructed. Based on the network, miR-223–3p was the hub node and the genes were significantly enriched in 15 GO terms of </span>biological processes and 14 KEGG pathways, including cAMP, PI3K-Akt, cGMP-PKG, </span>neurotrophin<span><span> signaling pathway, and </span>dopaminergic synapse. Then, miR-223–3p overexpressed AMSCs-derived exosomes were successfully extracted, and miR-223–3p was found to directly bind with </span></span><span><em>MAPK10</em></span>. <em>In vivo</em> experiments validated that AMSCs-derived exosomal miR-223–3p could promote wound healing, and up-regulated α-SMA, <span><em>CD31</em></span>, COL1A1, <em>COL2A1</em>, <em>COL3A1</em>, and down-regulated MAPK10, TNF-α, <em>IL-β</em>, and <em>IL-6</em>.</p></div><div><h3>Conclusions</h3><p>AMSC-derived exosomal miR-223–3p may accelerate wound healing by targeting MAPK10.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152102"},"PeriodicalIF":2.5,"publicationDate":"2023-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41187811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Saikosaponin-d regulates angiogenesis in idiopathic pulmonary fibrosis through angiopoietin/Tie-2 pathway","authors":"Yan Wu ,&nbsp;Jun Zhang ,&nbsp;Xintian Wang ,&nbsp;Yuncong Xu ,&nbsp;Jinxu Zheng","doi":"10.1016/j.acthis.2023.152100","DOIUrl":"10.1016/j.acthis.2023.152100","url":null,"abstract":"<div><h3>Objective</h3><p>Idiopathic pulmonary fibrosis (IPF) is considered as a chronic interstitial lung disease with underlying mechanism of IPF remaining unclear, while there are no definitive treatment options. In recent years, scientists have gradually paid attention to the influence of angiogenesis on IPF. Because IPF is a progressive with microvascular remodeling disorder, scientists have postulated that angiogenesis may also be one of the initiating and contributing factors of the disease. Bupleurum is a common natural Chinese herbal medicine with antibacterial, anti-inflammatory, anti-tumor and other pharmacological effects. As the most important active monomer of Bupleurum, Saikosaponin-d (SSd) is a new discovery with anti-pulmonary fibrosis (PF) activity. This study attempts to investigate the role of SSd in the interference of PF through regulation of angiogenesis in IPF through Angiopoietin (Angpt) /Tie receptor 2 (Tie2) pathway.</p></div><div><h3>Methods</h3><p>Randomly, we allocated C57BL/6 mice into four groups (n = 20 in each group). Afterwards, establishment of IPF model was accomplished via intratracheal administration of bleomycin (BLM, 5 mg/kg), while corresponding drug intervention was given accordingly. On 3rd, 7th, 14th and 28th days after modeling, we performed histopathological examination through staining. Meanwhile, immunohistochemistry (IHC) of PF and the expression of related factors were observed, while Ang/Tie2 pathway was assessed by ELISA with the effect of SSd on angiogenesis related proteins in IPF being explored with IHC and Western Blot technique.</p></div><div><h3>Results</h3><p>Our results showed that SSd could reduce inflammation and PF levels in lung tissue of experimental mice, while levels of angiogenesis-related factors, namely Tie-2, Ang-1 and ANGPT2 (Ang-2), fibrosis- associated factors like Alpha-smooth muscle actin (α-SMA), collagen-I and hydroxyproline in SSd and dexamethasone (DXM) mice were significantly reduced at each time point compared to BLM (p &lt; 0.01). Additionally, we discovered substantial decreased expressions of Ang-1, Ang-2, Tie-2, α-SMA and collagen-I at protein level in SSd and DXM mice at each time point compared to BLM (p &lt; 0.05). Besides, insignificant differences were observed between SSd and DXM groups (p &gt; 0.05).</p></div><div><h3>Conclusion</h3><p>This study has demonstrated that SSd could down-regulate the expression of ANG-1, Ang-2 and Tie2 in the Ang/Tie2 pathway, and may reduce lung inflammation and PF in BLM-induced mice via inhibition of angiogenesis.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152100"},"PeriodicalIF":2.5,"publicationDate":"2023-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41187812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibitory effect of telocyte-induced M1 macrophages on endometriosis: Targeting angiogenesis and invasion","authors":"Xiao-Jiao Wei ,&nbsp;Yue-lin Huang ,&nbsp;Tian-Quan Chen ,&nbsp;Xiao-Jun Yang","doi":"10.1016/j.acthis.2023.152099","DOIUrl":"10.1016/j.acthis.2023.152099","url":null,"abstract":"<div><h3>Purpose</h3><p>Telocytes (TCs), a novel type of stromal cells found in tissues, induce macrophage differentiation into classically activated macrophages (M1) types and enhance their phagocytic function. The purpose of this study was to investigate the inhibitory effects of TC-induced M1 macrophages on endometriosis (EMs).</p></div><div><h3>Methods</h3><p>mouse uterine primary TCs and endometrial stromal cells (ESCs) were isolated and identified using double immunofluorescence staining. For the <em>in vitro</em> study, ESCs were treated with TC-induced M1 macrophages, and the vascular endothelial growth factor (<em>VEGF</em>), matrix metalloproteinase 9 (<em>MMP9</em>), and nuclear factor kappa B (<em>NF-κb</em>) genes were identified by quantitative real-time PCR (qRT-PCR) or western blotting (WB). For the <em>in vivo</em> study, an EMs mouse model received TC-conditioned medium (TCM) <em>via</em> abdominal administration, and characterized the inhibitory effects on growth (lesion weight, volume, and pathology), tissue-resident macrophages differentiation by immunostaining, angiogenic capacity (CD31 and VEGF), invasive capacity (MMP9), and NF-κb expression within EMs lesions.</p></div><div><h3>Results</h3><p>immunofluorescent staining showed that uterine TCs expressed CD34+ and vimentin+, whereas ESCs expressed vimentin+ and cytokeratin-. At the cellular level, TC-induced M1 macrophages can significantly inhibit the expression of VEGF and MMP9 in ESCs through WB or qRT-PCR, possibly by suppressing the NF-κb pathway. The <em>in vivo</em> study showed that macrophages switch from the alternatively activated macrophages (M2) in untreated EMs lesions to the M1 subtype after TCM exposure. Thereby, TC-induced M1 macrophages contributed to the inhibition of EMs lesions. More importantly, this effect may be achieved by suppressing the expression of NF-κb to inhibit angiogenesis (CD31 and VEGF) and invasion (MMP9) in the tissue.</p></div><div><h3>Conclusion</h3><p>TC-induced M1 macrophages play a prevailing role in suppressing EMs by inhibiting angiogenic and invasive capacity through the NF-κb pathway, which provides a promising therapeutic approach for EMs.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152099"},"PeriodicalIF":2.5,"publicationDate":"2023-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41181745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transforming growth factor-beta superfamily regulates mesenchymal stem cell osteogenic differentiation: A microRNA linking","authors":"Juan F. Santibanez ,&nbsp;Cesar Echeverria ,&nbsp;Carola Millan ,&nbsp;Felipe Simon","doi":"10.1016/j.acthis.2023.152096","DOIUrl":"10.1016/j.acthis.2023.152096","url":null,"abstract":"<div><p><span>The ability to differentiate into cells of different lineages, such as bone cells, is the principal value of adult mesenchymal stem cells<span> (MSCs), which can be used with the final aim of regenerating damaged tissue. Due to its potential use and importance in regenerative medicine and </span></span>tissue engineering<span>, several questions have been raised regarding the molecular mechanisms of MSC differentiation. As one of the crucial mediators in organism development, the transforming growth factor-beta (TGF-β) superfamily directs MSCs’ commitment to selecting differentiation pathways. This review aims to give an overview of the current knowledge on the mechanisms of the TGF-β superfamily in MSCs bone differentiation, with additional insight into the mutual regulation of microRNAs and TGF-β in osteogenesis.</span></p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152096"},"PeriodicalIF":2.5,"publicationDate":"2023-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41181747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PGC1α deficiency reverses cholestasis-induced liver injury via attenuating hepatic inflammation and promoting bile duct remodeling","authors":"Dingwu Li ,&nbsp;Chenhui Ye ,&nbsp;Peihao Liu ,&nbsp;Ting Sun ,&nbsp;Yunsheng Qin ,&nbsp;Xingyong Wan","doi":"10.1016/j.acthis.2023.152097","DOIUrl":"10.1016/j.acthis.2023.152097","url":null,"abstract":"<div><h3>Objectives</h3><p><span>Cholestatic liver diseases are characterized by hepatocellular damage, </span>cholangiocyte<span><span><span> proliferation, and progressive fibrosis. Bile duct ligation (BDL) is widely used to resemble liver injuries induced by </span>cholestasis. Peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC1α) was reported to play a critical role in multiple biological responses. Nevertheless, whether PGC1α is involved in </span>bile acid metabolism and biliary disorders remains unclear. This study aimed to investigate the effect of PGC1α on hepatic responses after cholestatic injury.</span></p></div><div><h3>Materials and methods</h3><p>Wild-type mice were subjected to BDL or sham surgery for 14 days and human liver specimens from patients with primary biliary cholangitis<span> (PBC) were collected to detect the expression of PGC1α. Hepatic-specific PGC1α knockout mice<span> (HKO) were constructed and subjected to BDL, in which the effects of PGC1α on cholestatic liver injury were demonstrated by biochemical and histopathological assessments, immunoblotting<span>, and metabolomics.</span></span></span></p></div><div><h3>Results</h3><p><span>The expression of PGC1α was upregulated in the liver of PBC patients and murine models. Both in vivo and in vitro experiments supported the protective effects of PGC1α on cholestasis-induced hepatocyte injury. Infiltrated inflammatory cells<span><span> after BDL were decreased in HKO mice. Inhibited Wnt/β-Catenin pathway and enhanced Notch signaling promoted </span>transdifferentiation of </span></span>hepatic progenitor cells (HPC)/ hepatocytes into cholangiocytes, leading to the greater ductular reaction observed in the HKO mice. But bile acids metabolism and mitochondrial function were not affected due to hepatic PGC1α deficiency in cholestasis.</p></div><div><h3>Conclusions</h3><p>Hepatic-specific deletion of PGC1α regulated liver regeneration by promoting ductular reactions, thereby exerting protective effects against BDL-induced liver injury, which could be a new potential therapeutic target.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 8","pages":"Article 152097"},"PeriodicalIF":2.5,"publicationDate":"2023-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41181746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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 “Intracerebral injection of oil cyst content of human craniopharyngioma (oil machinery fluid) as a toxic model in the rat brain” [Acta Histochem. 116(3) (2014) 448–56]","authors":"Martha Lilia Tena-Suck ,&nbsp;Ma. Elena Hernández-Campos ,&nbsp;Alma Ortiz-Plata ,&nbsp;Citlaltepetl Salinas-Lara ,&nbsp;Ana Laura Colín-González ,&nbsp;Abel Santamaría","doi":"10.1016/j.acthis.2023.152089","DOIUrl":"10.1016/j.acthis.2023.152089","url":null,"abstract":"","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 7","pages":"Article 152089"},"PeriodicalIF":2.5,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10334880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Three-dimensional human germinal centers of different sizes in patients diagnosed with lymphadenitis show comparative constant relative volumes of B cells, T cells, follicular dendritic cells, and macrophages","authors":"Constantin Maximilian Schemel ,&nbsp;Patrick Wurzel ,&nbsp;Sonja Scharf ,&nbsp;Hendrik Schäfer ,&nbsp;Sylvia Hartmann ,&nbsp;Ina Koch ,&nbsp;Martin-Leo Hansmann","doi":"10.1016/j.acthis.2023.152075","DOIUrl":"10.1016/j.acthis.2023.152075","url":null,"abstract":"<div><p><span><span><span>Germinal centers (GCs) are some of the most important structures in the human immune system. As such, their cell types and functions have been thoroughly investigated. B cells, </span>T cells<span>, follicular dendritic cells (FDCs), and macrophages have widely been found to typically be aggregated in GCs. However, the amount of space occupied by each of these cell types has yet to be investigated. In this study, we conducted confocal laser-based 3D cell-volume quantification of typical GC cells under reactive conditions in </span></span>lymphadenitis<span><span> and investigated how volume proportions change during GC development. For this investigation, we used anti-CD3 (T cells), anti-CD20 and anti-Pax5 (B cells), anti-CD23 (FDCs), anti-CD68 (macrophages), and DAPI<span> (nuclear staining). We detected average proportions of about 11% CD3, 9% CD20, 6% </span></span>CD23<span>, and 2% CD68 in the largest possible regions of interest within GCs. Interestingly, these values remained steady relatively independent of GC size. The remarkably low B cell proportion can be attributed to technical constraints given the use of the CD20 antibody in 3D. Applying the B cell marker Pax5, we found that about 44% of the volume was occupied by B cells after extrapolating the volume of B </span></span></span>cell nuclei to that of whole B cells. We concluded that Pax5 is more suitable than anti-CD20 for 3D B cell quantification in GCs. The substantial unstained volume in GCs raises the question of whether other cell types fill these open spaces. Our 3D investigation enabled a unique morphological and volumetric evaluation of GC cells that balance their overall volumes in GCs.</p></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"125 7","pages":"Article 152075"},"PeriodicalIF":2.5,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9827136","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}