{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T07:48:36Z","timestamp":1787644116353,"version":"build-2736575974"},"reference-count":53,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2022,11,18]],"date-time":"2022-11-18T00:00:00Z","timestamp":1668729600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["22-74-10126"],"award-info":[{"award-number":["22-74-10126"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["21-74-20178"],"award-info":[{"award-number":["21-74-20178"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["075-15-2021-683"],"award-info":[{"award-number":["075-15-2021-683"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministry of Science and Higher Education of the Russian Federation","award":["22-74-10126"],"award-info":[{"award-number":["22-74-10126"]}]},{"name":"Ministry of Science and Higher Education of the Russian Federation","award":["21-74-20178"],"award-info":[{"award-number":["21-74-20178"]}]},{"name":"Ministry of Science and Higher Education of the Russian Federation","award":["075-15-2021-683"],"award-info":[{"award-number":["075-15-2021-683"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The mechanisms underlying the therapeutic potential of MSCs are the focus of intense research. We studied human MSCs isolated from desquamated endometrium (eMSCs), which, as previously shown, have high regenerative potential in various disease models. The aim was to evaluate the role of secreted VEGF in stimulating angiogenesis and maintaining eMSC viability and migration, which is important for improving the therapeutic properties of MSCs. We compared three eMSC cultures differing in the level of VEGF secretion: 3D spheroids, monolayer eMSCs, and monolayer eMSCs with VEGF knockdown. Spheroid eMSCs produced higher amounts of VEGF and had the strongest paracrine effect on HUVEC. eMSCs with VEGF knockdown did not stimulate angiogenesis. Monolayered eMSCs expressed VEGFR1, while spheroid eMSCs expressed both VEGFR1 and VEGFR2 receptors. The knockdown of VEGF caused a significant decrease in the viability and migration of eMSCs. eMSCs from 3D spheroids enhanced proliferation and migration in response to exogenous VEGF, in contrast to monolayered eMSCs. Our results suggest that the VEGF\u2013VEGFR1 loop appears to be autocrine-involved in maintaining the viability of eMSCs, and VEGFR2 expression enhances their response to exogenous VEGF, so the angiogenic potential of eMSC can be up- or downregulated by intrinsic VEGF signals.<\/jats:p>","DOI":"10.3390\/ijms232214324","type":"journal-article","created":{"date-parts":[[2022,11,21]],"date-time":"2022-11-21T02:52:21Z","timestamp":1668999141000},"page":"14324","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Paracrine and Autocrine Effects of VEGF Are Enhanced in Human eMSC Spheroids"],"prefix":"10.3390","volume":"23","author":[{"given":"Irina","family":"Kozhukharova","sequence":"first","affiliation":[{"name":"Institute of Cytology, Russian Academy of Sciences, 194064 Saint-Petersburg, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2528-046X","authenticated-orcid":false,"given":"Natalia","family":"Minkevich","sequence":"additional","affiliation":[{"name":"Institute of Cytology, Russian Academy of Sciences, 194064 Saint-Petersburg, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0814-9961","authenticated-orcid":false,"given":"Larisa","family":"Alekseenko","sequence":"additional","affiliation":[{"name":"Institute of Cytology, Russian Academy of Sciences, 194064 Saint-Petersburg, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alisa","family":"Domnina","sequence":"additional","affiliation":[{"name":"Institute of Cytology, Russian Academy of Sciences, 194064 Saint-Petersburg, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3128-4558","authenticated-orcid":false,"given":"Olga","family":"Lyublinskaya","sequence":"additional","affiliation":[{"name":"Institute of Cytology, Russian Academy of Sciences, 194064 Saint-Petersburg, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13045-021-01037-x","article-title":"Challenges and advances in clinical applications of mesenchymal stromal cells","volume":"14","author":"Zhou","year":"2021","journal-title":"J. Hematol. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1080\/14653240600855905","article-title":"Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement","volume":"8","author":"Dominici","year":"2006","journal-title":"Cytotherapy"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/j.cryobiol.2015.07.003","article-title":"Mesenchymal stromal cells derived from various tissues: Biological, clinical and cryopreservation aspects","volume":"71","author":"McGann","year":"2015","journal-title":"Cryobiology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1002\/stem.2922","article-title":"Concise review: Intercellular communication via organelle transfer in the biology and therapeutic applications of stem cells","volume":"37","author":"Krasnodembskaya","year":"2019","journal-title":"Stem Cells"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.cytogfr.2009.10.002","article-title":"Mechanisms involved in the therapeutic properties of mesenchymal stem cells","volume":"20","author":"Fontes","year":"2009","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1186\/s12967-014-0260-8","article-title":"A review of therapeutic effects of mesenchymal stem cell secretions and induction of secretory modification by different culture methods","volume":"12","author":"Madrigal","year":"2014","journal-title":"J. Transl. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1093\/bmb\/ldt031","article-title":"The angiogenic properties of mesenchymalstem\/stromal cells and their therapeutic potential","volume":"108","author":"Watt","year":"2013","journal-title":"Br. Med. Bull."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Harrell, C.R., Fellabaum, C., Jovicic, N., Djonov, V., Arsenijevic, N., and Volarevic, V. (2019). Molecular mechanisms responsible for the rapeutic potential of mesenchymal stem cell-derived secretome. Cells, 8.","DOI":"10.3390\/cells8050467"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"107","DOI":"10.19104\/jbtr.2014.107","article-title":"Mesenchymal stem cell paracrine factors in vascular repair and regeneration","volume":"1","author":"Pankajakshan","year":"2014","journal-title":"J. Biomed. Technol. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/386671a0","article-title":"Mechanisms of angiogenesis","volume":"386","author":"Risau","year":"1997","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2235","DOI":"10.1172\/JCI116450","article-title":"Patterns of expression of vascular endothelial growth factor (VEGF) and VEGF receptors in mice suggest a role in hormonally regulated angiogenesis","volume":"91","author":"Shweiki","year":"1993","journal-title":"J. Clin. Investig."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1042\/BJ20110301","article-title":"Signal transduction by vascular endothelial growth factor receptors","volume":"437","author":"Koch","year":"2011","journal-title":"Biochem. J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"26988","DOI":"10.1016\/S0021-9258(18)47116-5","article-title":"Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor","volume":"269","author":"Waltenberger","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1096\/fasebj.13.1.9","article-title":"Vascular endothelial growth factor (VEGF) and its receptors","volume":"131","author":"Neufeld","year":"1999","journal-title":"FASEB J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41540-017-0037-9","article-title":"VEGFR1 promotes cell migration and proliferation through PLC\u03b3 and PI3K pathways","volume":"4","author":"Weddell","year":"2018","journal-title":"NPJ Syst. Biol. Appl."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"13313","DOI":"10.1074\/jbc.273.21.13313","article-title":"Vascular endothelial growth factor induces expression of the antiapoptotic proteins Bcl-2 and A1 in vascular endothelial cells","volume":"273","author":"Gerber","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1016\/j.cell.2007.06.054","article-title":"Autocrine VEGF signaling is required for vascular homeostasis","volume":"130","author":"Lee","year":"2007","journal-title":"Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2236","DOI":"10.1242\/jcs.163774","article-title":"Autocrine VEGF maintains endothelial survival through regulation of metabolism and autophagy","volume":"128","author":"Domigan","year":"2015","journal-title":"Cell Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1161\/01.RES.0000118601.37875.AC","article-title":"Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms","volume":"94","author":"Kinnaird","year":"2004","journal-title":"Circ. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1292","DOI":"10.1161\/01.CIR.0000121425.42966.F1","article-title":"Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells","volume":"109","author":"Rehman","year":"2004","journal-title":"Circulation"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1134\/S1990519X20030050","article-title":"Characterization of cultured mesenchymal stromal cells established from human chorion","volume":"14","author":"Shilina","year":"2020","journal-title":"Cell Tissue Biol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Santos, R.d.A., Asensi, K.D., de Barros, J.H.O., de Menezes, R.C.S., Cordeiro, I.R., Neto, J.M.d.B., Kasai-Brunswick, T.H., and Goldenberg, R.C.d.S. (2020). Intrinsic angiogenic potential and migration capacity of human mesenchymal stromal cells derived from menstrual blood and bone marrow. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21249563"},{"key":"ref_23","first-page":"1667","article-title":"VEGF secreted by mesenchymal stem cells mediates the differentiation of endothelial progenitor cells into endothelial cells via paracrine mechanisms","volume":"17","author":"Ge","year":"2017","journal-title":"Mol. Med. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"15554","DOI":"10.1074\/jbc.M602003200","article-title":"Expression of vascular endothelial growth factor receptor 1 in bone marrow-derived mesenchymal cells is dependent on hypoxia-inducible factor 1","volume":"281","author":"Okuyama","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1002\/jcb.20462","article-title":"Vascular endothelial growth factor (VEGF-A) expression in human mesenchymal stem cells: Autocrine and paracrine role on osteoblastic and endothelial differentiation","volume":"95","author":"Mayer","year":"2005","journal-title":"J. Cell. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3101","DOI":"10.1172\/JCI61209","article-title":"Intracellular VEGF regulates the balance between osteoblast and adipocyte differentiation","volume":"122","author":"Liu","year":"2012","journal-title":"J. Clin. Investig."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.ejvs.2017.10.012","article-title":"Efficient differentiation of bone marrow mesenchymal stem cells into endothelial cells in vitro","volume":"55","author":"Wang","year":"2018","journal-title":"Eur. J. Vasc. Endovasc. Surg."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1634\/stemcells.22-3-377","article-title":"Mesenchymal stem cells can be differentiated into endothelial cells in vitro","volume":"22","author":"Oswald","year":"2004","journal-title":"Stem Cells"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3845","DOI":"10.1210\/en.2008-1207","article-title":"Vascular endothelial growth factor receptor 2 (VEGFR-2) functions to promote uterine decidual angiogenesis during early pregnancy in the mouse","volume":"150","author":"Douglas","year":"2009","journal-title":"Endocrinology"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Guo, X., Yi, H., Li, T.C., Wang, Y., Wang, H., and Chen, X. (2021). Role of vascular endothelial growth factor (VEGF) in human embryo implantation: Clinical Implications. Biomolecules, 11.","DOI":"10.3390\/biom11020253"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1186\/s13287-018-0801-9","article-title":"Human mesenchymal stem cells in spheroids improve fertility in model animals with damaged endometrium","volume":"9","author":"Domnina","year":"2018","journal-title":"Stem Cell Res. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/j.stem.2010.11.015","article-title":"Reprogramming of human primary somatic cells by Oct4 and chemical compounds","volume":"7","author":"Zhu","year":"2010","journal-title":"Cell Stem Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1101\/gad.1388806","article-title":"Smad3-dependent nuclear translocation of beta-catenin is required for TGF-beta1-induced proliferation of bone marrow-derived adult human mesenchymal stem cells","volume":"20","author":"Jian","year":"2006","journal-title":"Genes Dev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1038\/sj.bjc.6604508","article-title":"VEGF expression by mesenchymal stem cells contributes to angiogenesis in pancreatic carcinoma","volume":"99","author":"Beckermann","year":"2008","journal-title":"Br. J. Cancer"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1016\/S0002-9440(10)64964-4","article-title":"Hypoxia-induced vascular endothelial growth factor expression precedes neovascularization after cerebral ischemia","volume":"156","author":"Marti","year":"2000","journal-title":"Am. J. Pathol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"473","DOI":"10.3389\/fcell.2020.00473","article-title":"Three-dimensional compaction switches stress response programs and enhances therapeutic efficacy of endometrial mesenchymal stem\/stromal cells","volume":"8","author":"Domnina","year":"2020","journal-title":"Front. Cell. Dev. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2363","DOI":"10.1634\/stemcells.2006-0686","article-title":"Angiogenic effects of human multipotent stromal cell conditioned medium activate the PI3K-Akt pathway in hypoxic endothelial cells to inhibit apoptosis, increase survival, and stimulate angiogenesis","volume":"25","author":"Hung","year":"2007","journal-title":"Stem Cells"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1761","DOI":"10.1634\/stemcells.2007-0022","article-title":"Mesenchymal stem cells support migration, extracellular matrix invasion, proliferation, and survival of endothelial cells in vitro","volume":"25","author":"Potapova","year":"2007","journal-title":"Stem Cells"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.actbio.2019.09.022","article-title":"Characterizing the impact of 2D and 3D culture conditions on the therapeutic effects of human mesenchymal stem cell secretome on corneal wound healing in vitro and ex vivo","volume":"99","author":"Carter","year":"2019","journal-title":"Acta Biomater."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1172\/JCI69244","article-title":"Hypoxia-inducible factor\u2013dependent breast cancer\u2013mesenchymal stem cell bidirectional signaling promotes metastasis","volume":"123","author":"Chaturvedi","year":"2013","journal-title":"J. Clin. Investig."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1794","DOI":"10.1002\/bit.24827","article-title":"Hypoxia induces osteogenic\/angiogenic responses of bone marrow-derived mesenchymal stromal cells seeded on bone-derived scaffolds via ERK1\/2 and p38 pathways","volume":"110","author":"Zhou","year":"2013","journal-title":"Biotechnol. Bioeng."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"8703","DOI":"10.1111\/jcmm.15502","article-title":"Growth differentiation factor 11 promotes differentiation of MSCs into endothelial-like cells for angiogenesis","volume":"24","author":"Zhang","year":"2020","journal-title":"J. Cell. Mol. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1182\/blood.V97.3.785","article-title":"Flt-1, vascular endothelial growth factor receptor 1, is a novel cell surface marker for the lineage of monocyte-macrophages in humans","volume":"97","author":"Sawano","year":"2001","journal-title":"Blood"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2138","DOI":"10.1038\/sj.onc.1203533","article-title":"Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells","volume":"19","author":"Kanno","year":"2000","journal-title":"Oncogene"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"24599","DOI":"10.1021\/acsami.0c05681","article-title":"Exogenous signaling molecules released from aptamer-functionalized hydrogels promote the survival of mesenchymal stem cell spheroids","volume":"12","author":"Zhao","year":"2020","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1161\/CIRCRESAHA.108.174128","article-title":"Vascular endothelial growth factor receptor-1 regulates postnatal angiogenesis through inhibition of the excessive activation of AKT","volume":"103","author":"Nishi","year":"2008","journal-title":"Circ. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"38527","DOI":"10.1074\/jbc.M104536200","article-title":"Synergistic cooperation between hypoxia and transforming growth factor-beta pathways on human vascular endothelial growth factor gene expression","volume":"276","author":"Botella","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e1317","DOI":"10.1038\/cddis.2014.282","article-title":"TGF-\u03b2\/Smad3 inhibit vascular smooth muscle cell apoptosis through an autocrine signaling mechanism involving VEGF-A","volume":"5","author":"Shi","year":"2014","journal-title":"Cell Death Dis."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2671","DOI":"10.1002\/jcp.24983","article-title":"Cross-talk between VEGF and BMP-6 pathways accelerates osteogenic differentiation of human adipose-derived stem cells","volume":"230","author":"Li","year":"2015","journal-title":"J. Cell. Physiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1705","DOI":"10.1089\/ten.tea.2014.0314","article-title":"Compaction, fusion, and functional activation of three-dimensional human mesenchymal stem cell aggregate","volume":"21","author":"Tsai","year":"2015","journal-title":"Tissue Eng. Part A"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"584","DOI":"10.5966\/sctm.2013-0007","article-title":"Short-term spheroid formation enhances the regenerative capacity of adipose-derived stem cells by promoting stemness, angiogenesis, and chemotaxis","volume":"2","author":"Cheng","year":"2013","journal-title":"Stem Cells Transl. Med."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"13100","DOI":"10.1074\/jbc.M800184200","article-title":"Culturing of human mesenchymal stem cells as three-dimensional aggregates induces functional expression of CXCR4 that regulates adhesion to endothelial cells","volume":"283","author":"Potapova","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_53","first-page":"919","article-title":"Multipotent mesenchymal stem cells of desquamated endometrium: Isolation, characterization, and application as a feeder layer for maintenance of human embryonic stem cells","volume":"53","author":"Zemelko","year":"2011","journal-title":"Tsitologiia"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/22\/14324\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:21:16Z","timestamp":1760145676000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/22\/14324"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,18]]},"references-count":53,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["ijms232214324"],"URL":"https:\/\/doi.org\/10.3390\/ijms232214324","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,18]]}}}