Cytotechnology最新文献

{"title":"Docetaxel treatment together with CTLA-4 knockdown enhances reduction of cell viability and amplifies apoptosis stimulation of MCF-7 breast cancer cells.","authors":"Negar Hosseinkhani, Shiva Alipour, Amir Ghaffari Jolfayi, Leili Aghebati-Maleki, Elham Baghbani, Nazila Alizadeh, Vahid Khaze, Behzad Baradaran","doi":"10.1007/s10616-024-00677-4","DOIUrl":"https://doi.org/10.1007/s10616-024-00677-4","url":null,"abstract":"<p><p>Breast cancer is the most frequent cancer in women with a 20% mortality rate. The fate of patients suffering from breast cancer can be influenced by immune cells and tumor cells interaction in the tumor microenvironment (TME). Immune checkpoints such as Cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) are regulators of the immune system and defend normal tissues from immune cell attacks but they can be expressed in breast cancer tissue and facilitate immune evasion of tumoral cells. Based on this, here we studied the role of CTLA-4 silencing by specific siRNA in MCF-7 breast cancer cell line together with Docetaxel treatment which is one of the robust chemotherapy agents to demonstrate the significance of combining chemotherapy with efficient targeted therapy in tumor regression. The MCF-7 breast cancer cell line was transfected with CTLA-4-siRNA through the electroporation method, then received an appropriate dose of Docetaxel determined by MTT assay. Flow cytometry was utilized to investigate the consequence of simultaneous CTLA-4 gene silencing and Docetaxel treatment on the apoptosis and cell cycle of MCF-7 cells. The expression levels of Bax and Bcl-2 were also investigated using quantitative real-time PCR. Compared to control groups, CTLA-4-suppressed and Docetaxel-treated cells became more susceptible to apoptosis and cell cycle arrest at the G2-M phase. The additive effect of CTLA-4 knockdown together with Docetaxel treatment significantly downregulated BCL-2 level and upregulated BAX expression. Our findings support the idea that combining chemotherapy such as Docetaxel with efficient targeted therapy against inhibitory immune checkpoints can be a promising strategy in cancer treatment.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"19"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11638433/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142827603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"IBS008738, a TAZ activator, facilitates muscle repair and inhibits muscle injury in a mouse model of sport-induced injury.","authors":"Yiming Wang, Datian Liu, Sining Wang, Yiliang Li, Guanming Liu","doi":"10.1007/s10616-024-00667-6","DOIUrl":"10.1007/s10616-024-00667-6","url":null,"abstract":"<p><p>High-intensity exercise can cause excessive generation of ROS and induce oxidative stress injury in the body, which is a major reason accounting for muscle damage following exercise. The previous study demonstrated that IBS008738, the activator of TZA, was able to enhance myogenesis in mouse myogenic C2C12 cells, prevent dexamethasone-induced muscle atrophy, and facilitate muscle repair in cardiotoxin-induced muscle injury. Accordingly, our study was designed to probe into the potential role of IBS008738 in muscle damage in mouse models induced by high-intensity exercise. Mice were first administrated with IBS008738, and then subjected to high-intensity eccentric exercise to induce muscle damage after 24 h. During the experiment, mouse weight change and food take were recorded. At the end of the experiment, blood samples were collected through cardiac puncture and centrifugated. Serum levels of blood urea nitrogen (BUN), creatinine, glucose, lactate dehydrogenase (LDH), creatinine kinase (CK), and C-related protein were evaluated using an autoanalyzer. After mice were sacrificed, the gastrocnemius muscles were dissected for DCFH-DA assay of ROS generation, thiobarbituric acid-reactive substances (TBARS) assay of MDA content, hematoxylin-eosin (H&E) staining of histological examination, and western blotting analysis of Akt/mTOR/S6K1 signaling expression. IBS008738 and/or exercise exert significant effects on mouse weight and food take. High-intensity exercise markedly increased ROS generation and lipid peroxidation, upregulated serum levels of CK, LDH, and C-related protein, ameliorated muscle histological damage, and reduced TAZ, phosphorylated (p)-Akt, p-mTOR, and p-S6K1 protein levels in mice. However, IBS008738 administration reversed the above changes induced by high-intensity exercise in mice. IBS008738 alleviates oxidative stress and muscle damage in mice after high-intensity exercise by activating TAZ and the Akt/mTOR/S6K1 signaling pathway.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-024-00667-6.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"2"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11576750/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of cell lysis for improved understanding between the shake tube and stirred tank reactor perfusion CHO cell cultures.","authors":"Weijian Zhang, Qingyuan Ran, Yang Zhou, Liang Zhao, Qian Ye, Wen-Song Tan","doi":"10.1007/s10616-024-00662-x","DOIUrl":"https://doi.org/10.1007/s10616-024-00662-x","url":null,"abstract":"<p><p>Shake tubes (ST) are widely employed to assist the development of the stirred tank reactor (STR) perfusion cell culture. However, cell lysis may be frequently underrestimated and lead to culture performance discrepency between these systems, rendering the ST model ineffective in designing the STR perfusion cultures. In this study, perfusion culture performance bewteen the STR and ST was investigated under various conditions with the analysis of cell lysis. Comparable performance was observed bewteen the two systems at low perfusion rates ( <math><mi>D</mi></math> ≤1.0 VVD), except that the specific productivity ( <math><msub><mi>q</mi> <mi>P</mi></msub> </math> ) of the STR was decreased at <math><mi>D</mi></math> =0.5 VVD, which was related to product degradation by cell lysis. In contrast, significant differences in cell maintenance, metabolism, and <math><msub><mi>q</mi> <mi>P</mi></msub> </math> were found at <math><mi>D</mi></math> =2.0 VVD. By the analysis of the authentic cell growth and death kinetics, it was found that cell growth arrest, potentially due to the limited availability of oxygen, led to the stable cell maintenance at VCD≈90 × 10⁶ cells/ml and altered cellular metabolism for the ST, while the continuous decline of VCD and <math><msub><mi>q</mi> <mi>P</mi></msub> </math> in the STR were related to excessive cell death, subsequently ascribed to the harmful hydrodynamic stress conditions. We further demonstrated that cell lysis accounted for 57.62-76.29% of the total generated biomass in both the reactors and significantly impacted the estimation of process descriptors crucial for understanding the true cellular states. With cell lysis in sight, cell performance can therefore be accurately described and this knowledge can be further leveraged to expedite process development for the perfusion cell culture processes.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"7"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11602925/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aging and cell expansion enhance microRNA diversity in small extracellular vesicles produced from human adipose-derived stem cells.","authors":"Toshiya Tsubaki, Ryota Chijimatsu, Taiga Takeda, Maki Abe, Takahiro Ochiya, Shinsaku Tsuji, Keita Inoue, Tokio Matsuzaki, Yasuhide Iwanaga, Yasunori Omata, Sakae Tanaka, Taku Saito","doi":"10.1007/s10616-024-00675-6","DOIUrl":"10.1007/s10616-024-00675-6","url":null,"abstract":"<p><p>Adipose-derived stem cells (ASCs) and their small extracellular vesicles (sEVs) hold significant potential for regenerative medicine due to their tissue repair capabilities. The microRNA (miRNA) content in sEVs varies depending on ASC status; however, the effects of aging and cell passage on miRNA profiles remain unclear. In this study, we examined the effects of donor age and cell expansion on ASC characteristics and transcriptome using ASCs obtained from three young and three old donors. Cell expansion significantly impaired stem cell properties, notably reducing proliferation and differentiation capacities. In contrast, donor age had minimal effects on ASCs. RNA sequencing (RNA-seq) revealed differences in gene expression related to stemness, phagocytosis, and metabolic processes influenced by cell expansion. To investigate miRNA variability, we performed small RNA-seq on sEVs collected from ASCs of all six donors. The miRNA profiles were influenced by donor age and cell passage. Interestingly, functional enrichment analysis indicated that advanced donor age and increased cell passage may enhance the production of miRNAs associated with organ development through various pathways. These findings suggest that donor age and cell expansion differentially influence ASC characteristics and sEV miRNA content, highlighting the need for disease-specific conditioning of ASCs to optimize the therapeutic effects of sEVs in clinical applications.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-024-00675-6.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"15"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11631832/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142812659","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of a modified lactate dehydrogenase assay to evaluate the viability of cells cultured on 3D scaffolds when commonly used assays fail.","authors":"Zhanna K Nazarkina, Alena O Stepanova, Tatyana A Savostyanova, Pavel P Laktionov","doi":"10.1007/s10616-024-00670-x","DOIUrl":"https://doi.org/10.1007/s10616-024-00670-x","url":null,"abstract":"<p><p>The development of new compounds or materials for medicine suggests a study of their cytotoxicity. The effect of a material on cell viability can be evaluated by several methods based on DNA content, DNA synthesis, plasma membrane integrity, cellular enzyme activity, cellular reducing potential, and ATP level. Sometimes it is impossible to apply widely used commercially available reagents, e.g., when cells are cultured on materials, that interfere with the chemicals used or resulting in the course of enzymatic reaction. Here, we offer a method for monitoring the viability of cells proliferating on different supports in vitro. The method is based on the measurement of lactate dehydrogenase (LDH) activity in cellular lysates. After cells were lysed in 1% Nonidet P40 and supernatants were transferred into fresh tubes, LDH activity was measured in the supernatants using colorimetric method. The usefulness of the test was studied using human cervical adenocarcinoma HeLa cells and human gingival fibroblasts cultivated on different materials, including activated carbon-loaded scaffolds. The comparison with widely used AlamarBlue® assay confirms the LDH-based method as an appropriate alternative for measuring the living cell number in vitro in a quick, simple, and cost-effective manner when widespread methods for the evaluation of cell viability could not be used.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"9"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11604983/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synovial fluid mesenchymal stem cell-derived microRNA-127-5p can modulate transforming growth factor-β signaling after in vitro chondrogenic induction.","authors":"Tugba Semerci Sevimli, Ulukan Inan, Emilia Qomi Ekenel, Cemre Ozgul, Cem Ozgur Danaci, Sevval Cetinkaya, Zarifa Ahmadova","doi":"10.1007/s10616-024-00660-z","DOIUrl":"https://doi.org/10.1007/s10616-024-00660-z","url":null,"abstract":"<p><p>MicroRNA profiling in human cartilage is necessary for chondrogenesis. The study aimed to compare microRNA 127-5p (miR-127-5p) and TGF-β signaling pathway gene expressions of human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and synovial fluid-derived stem cells (hSF-MSCs) after induced chondrogenesis. MSCs induced into chondrogenic differentiation. Alcian Blue and Safranin O staining were performed to determine chondrogenic differentiation. The RT-qPCR determined the expression levels of miR-127-5p and TGF-β signaling pathway genes. miR-127-5p expression was significantly higher in chondrogenic differentiated hSF-MSCs (dhSF-MSCs) (p < 0.05). <i>TGF-β</i>, <i>SMAD2</i>, and <i>SMAD3</i> expressions were substantially higher in dhSF-MSCs (all p < 0.001), while <i>SMAD4</i>, and <i>ACAN</i> expressions were downregulated (all p < 0.001). No difference was detected between <i>COL1A2</i> expression levels. This study suggests that miR-127-5p derived from hSF-MSCs may regulate chondrogenesis, thereby inducing the <i>TGF-β</i> pathway activation, and also presents, for the first time, a comparative analysis of the expression of miR-127-5p and the TGF-β signaling pathway genes of hSF-MSCs and hAT-MSCs concerning differences in chondrogenic potential.</p><p><strong>Graphical abstract: </strong></p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"8"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11602905/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of scaffold manufacturing conditions for 3-dimensional culture of myogenic cell line derived from black sea bream (<i>Acanthopagrus schlegelii</i>).","authors":"Ye-Eun Lee, Eun Soo Jeong, Young-Mog Kim, Seung Pyo Gong","doi":"10.1007/s10616-024-00676-5","DOIUrl":"10.1007/s10616-024-00676-5","url":null,"abstract":"<p><p>Culturing fish myogenic cells in vitro holds significant potential to revolutionize aquaculture practices and support sustainable food production. However, advancement in in vitro culture technologies for skeletal muscle-derived myogenic cells have predominantly focused on mammals, with limited studies on fish. Scaffold-based three-dimensional (3D) culture systems for fish myogenic cells remain underexplored, highlighting a critical research gap compared to mammalian systems. This study evaluated the effects of scaffold composition and manufacturing methods on cellular growth in the 3D culture of black sea bream (<i>Acanthopagrus schlegelii</i>) myogenic cells. Scaffolds were manufactured using three natural polymers: black sea bream-derived extracellular matrix (ECM), sodium alginate, and gelatin. Two scaffold types were tested: \"cell-laden scaffolds\" prepared by mixing cells into the pre-scaffold solution followed by gelation, and \"cell-seeding scaffolds\" produced by freezing, gelation, and lyophilization before cell inoculation. Scaffold characteristics, including pore size, porosity, swelling ratio, and degradation rate, were assessed. Cell-seeding scaffolds exhibited relatively larger pore size, higher porosity, and higher degradation rate, while cell-laden scaffolds had higher swelling ratios. When black sea bream myogenic cells were cultured in these scaffolds, cell-seeding scaffolds supported cellular growth, particularly when composed of 3% sodium alginate and 4% gelatin with any concentration of ECM. In contrast, cell-laden scaffolds did not support cellular growth regardless of their composition. These findings provide fundamental insights for optimizing scaffold properties to develop more optimized conditions for 3D culture of fish muscle lineage cells.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"18"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11635079/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142827626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional improvements in β-lactoglobulin by conjugation with high methoxy pectin by the Maillard reaction.","authors":"Koya Shibata, Tomomi Odashima, Taku Harada, Yuichiro Ohno, Tadashi Yoshida, Makoto Hattori","doi":"10.1007/s10616-024-00665-8","DOIUrl":"10.1007/s10616-024-00665-8","url":null,"abstract":"<p><p>β-lactoglobulin (BLG), a major protein in whey, was conjugated with high methoxy pectin (HMP) by the Maillard reaction to improve emulsifying property and reduce allergenicity of BLG. The Maillard reaction was carried out at a weight ratio BLG: HMP = 1:4.15 and 1:6 at 60 °C at a relative humidity of 79% for 5 days. Crude conjugates were purified by dialysis and anion exchange chromatography. These two conjugates were named Conj. LS and Conj. HS, respectively. The results of SDS-PAGE showed that conjugates of various molecular weights were generated, and the weight ratios of protein to saccharide of Conj. LS and Conj. HS were 1:2.15 and 1:6.44 respectively, and the degree of reduction of free amino groups was 6.1 and 6.7 respectively. Emulsifying property of BLG was significantly improved by both conjugation. Both conjugates showed excellent emulsifying property in the acidic pH and in the presence of NaCl. Conjugation with HMP significantly reduced immunogenicity, which was more pronounced in conj. HS. Conjugation with HMP was considered to be an effective method to improve the functionality of BLG. Conjugation method used in this study is a safe method that is considered to be very valuable for food processing.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"3"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11576664/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Animal origins free products in cell culture media: a new frontier.","authors":"Mahsa Golshan, Hengameh Dortaj, Mehrdad Rajabi, Zeinab Omidi, Mehdi Golshan, Majid Pourentezari, Ali Rajabi","doi":"10.1007/s10616-024-00666-7","DOIUrl":"10.1007/s10616-024-00666-7","url":null,"abstract":"<p><p>Despite the importance of finding replacements for fetal bovine serum (FBS), very few studies have focused on this subject. Historically, the use of animals and their derivatives in growth, reproduction, and physiological studies has raised several concerns. The supplementation of culture media with FBS, also known as fetal calf serum, continues to be widespread, despite its limitations in quality, reproducibility, and implications for animal welfare. Moreover, the presence of counterfeit and illegal products can adversely affect cell cultures and treatments, prompting the search for alternative solutions. To reduce reliance on FBS, various substitutes have been introduced, such as plant-derived proteins, bovine eye fluid, sericin protein, human platelet lysate, and inactivated coelomic fluid, which can provide roles similar to that of FBS. Therefore, it is essential to develop serum-free and animal supplement-free environments suitable for therapeutic and clinical applications, tailored to the specific needs of different cell types. Among the alternatives, plant-based options have gained attention as sustainable and ethical solutions. These include plant-derived peptones from sources like soy and wheat, which are rich in amino acids and peptides essential for mammalian cell growth, as well as plant protein hydrolysates from beans and peas that serve as sources of amino acids and growth factors. Plant extracts, especially from soy and various seeds, contain necessary proteins and growth factors, while phytohormones such as cytokinins and plant polysaccharides can help regulate cell growth. While these alternatives offer benefits like reduced costs and lower risks of disease transmission, further research is necessary to refine and align them with the specific requirements of diverse cell types.</p><p><strong>Graphical abstract: </strong></p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"12"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11625046/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142799186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of the effects of different hypoxia mimetic agents on the secretome contents of conditioned medium obtained from mesenchymal stem/stromal cells cultured in 2 or 3-dimensional cell culture systems.","authors":"Serbay Ozkan, Basak Isildar, Meral Koyuturk","doi":"10.1007/s10616-024-00659-6","DOIUrl":"10.1007/s10616-024-00659-6","url":null,"abstract":"<p><p>Paracrine factors secreted by mesenchymal stem/stromal cells (MSCs) have been demonstrated to have significant therapeutic potential. The secretome profiles of MSCs variate depending on culture conditions. Generally, the effects of a single preconditioning strategy on secretome profiles of MSCs were investigated. However, until now, there has been no study examining the combinatory effects of different preconditioning strategies in a comparative manner. This study aimed to evaluate the secretome contents of conditioned media obtained from human umbilical cord-derived MSCs cultured in 2- or 3-dimensional (D) culture systems preconditioned with deferoxamine (DFS) or dimethyloxalylglycine (DMOG). Immunocytochemical analysis showed that MSCs preconditioned with DFS or DMOG have increased nuclear hypoxia-inducible factor-1α expression. Transmission electron microscopic analysis showed that cells preconditioned with DFS or DMOG have increased autophagic vesicles, which could be attributed to altered energy metabolism under hypoxic conditions. It was revealed that hypoxia-mimetic agents added to the 2D-, or 3D-culture environment raised total protein concentrations per cell along with vascular endothelial growth factor. The concentrations of glial cell-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) were differentially regulated in 2D-, and 3D-culture system, that the secretions of GDNF and NGF per cell were more prominent in 3D- and 2D-culture systems, respectively. These findings indicate that hypoxic conditions alone significantly elevate total protein concentrations, while the contribution of the 3D environment is more modest than initially anticipated. However, concentrations of secreted growth factors may be affected differently depending on the topography of the culture condition and the types of hypoxia mimetic agents.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 1","pages":"11"},"PeriodicalIF":2.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11625095/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142799499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}