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However, the specific conditions under which the influence of fibroblasts becomes important in modulating angiogenic formation remain to be determined. Here, we extend a previously established microfluidic co-culture system to enable precise manipulation of the density of human lung fibroblasts (HLF) and their distance to human umbilical vein endothelial cells (HUVECs) in a wide range within fibrin hydrogels. Using live cell imaging and image analysis, we quantify the effects of fibroblast density, HLF- HUVEC cell\u2013cell distance, and culturing time on cell migration, morphology, microvasculature, and endothelial sprouting. We find that, within the system and parameter range explored here, for fibroblasts to have a significant positive impact in the angiogenesis of endothelial cells, the fibroblast density should be no less than ~3\u00d710\n                    <jats:sup>6<\/jats:sup>\n                    cells\/ml and the cell\u2013cell distance should be no greater than ~2\u00a0mm. The experimental findings can be captured by a minimal model accounting for the concentration profile of pro-angiogenic factors secreted by fibroblasts. Our results provide insights into the remodeling of vasculature and the guidance for engineering vascularized tissue mimics.\n                  <\/jats:p>","DOI":"10.1007\/s10404-026-02939-3","type":"journal-article","created":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T05:14:52Z","timestamp":1792041292000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Spatial distribution and density of fibroblasts determine angiogenic response of endothelial cells"],"prefix":"10.1007","volume":"30","author":[{"given":"Pu","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melur K.","family":"Ramasubramanian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li-Heng","family":"Cai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"key":"2939_CR1","doi-asserted-by":"publisher","first-page":"3261","DOI":"10.1089\/ten.tea.2009.0708","volume":"16","author":"KA Ahmann","year":"2010","unstructured":"Ahmann KA, Weinbaum JS, Johnson SL, Tranquillo RT (2010) Fibrin degradation enhances vascular smooth muscle cell proliferation and matrix deposition in fibrin-based tissue constructs fabricated in vitro. 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