{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,10]],"date-time":"2026-05-10T08:58:12Z","timestamp":1778403492251,"version":"3.51.4"},"reference-count":34,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T00:00:00Z","timestamp":1771459200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32101114"],"award-info":[{"award-number":["32101114"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81871450"],"award-info":[{"award-number":["81871450"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Funding for Open Projects of the Key Laboratory of Acupuncture-Moxibustion and Tuina Intel-ligent Equipment of Chongqing Administration of Traditional Chinese Medicine","award":["ZJTNZDSYS01-01"],"award-info":[{"award-number":["ZJTNZDSYS01-01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Microfluidics-based preparation methods for cell-laden hydrogel microspheres are well-suited for large-scale comparative analysis of single or few cells. However, in existing studies, the preparation of cell-laden hydrogel microspheres and the cell culture process are typically separated, requiring the fabricated microspheres to be eluted and transferred from the preparation device to cell culture dishes or plates for cultivation. This transfer process can easily compromise sterility, while conventional cell culture methods consume more reagents and cause microsphere stacking, hindering single-cell observation and analysis. To address these issues, this paper presents an integrated microfluidic chip that sequentially enables droplet generation with cell encapsulation, gel droplet solidification, hydrogel microsphere trapping, and microsphere-based cell culture and analysis, facilitating the cultivation and observation of single or small numbers of cells. Integrating cell-laden microsphere preparation and 3D cell culture within a sealed chip structure reduces contamination risks associated with cell transfer, enables automation of multiple cell analysis workflows, and minimizes reagent and sample consumption. Using polydimethylsiloxane (PDMS) with good gas permeability and processability as the chip material, biocompatible fluorinated oil was selected as the oil phase for microsphere preparation. A mild sodium alginate-calcium ion gelation system was employed, where calcium ions were released under acidic conditions after droplet generation to trigger solidification, yielding uniform hydrogel microspheres. Under optimized conditions, the single-cell encapsulation efficiency for test samples of human myeloid leukemia cells (K562) was 33.8% \u00b1 1.8%, with a size uniformity coefficient of variation (CV) reaching 3.85%. Cells encapsulated within hydrogel microspheres were cultured in 286 on-chip independent cell culture chambers, achieving &gt;95% viability after 24 h.<\/jats:p>","DOI":"10.3390\/bios16020126","type":"journal-article","created":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T08:50:03Z","timestamp":1771577403000},"page":"126","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Integrated Microfluidic Chip Enabling Preparation and Immobilization of Cell-Laden Microspheres, and Microsphere-Based Cell Culture and Analysis"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3678-439X","authenticated-orcid":false,"given":"Qiongyao","family":"Mou","sequence":"first","affiliation":[{"name":"Key Laboratory of Acupuncture-Moxibustion and Tuina Intelligent Equipment of Chongqing Administration of Traditional Chinese Medicine, Chongqing Three Gorges Medical College, Chongqing 404120, China"},{"name":"Research Center of Oral Materials and Technology, Department of Medical Technology, Chongqing Three Gorges Medical College, Chongqing 404120, China"},{"name":"Key Laboratory of Biorheological Science and Technology, Ministry of Education and Bioengineering College, Chongqing University, Chongqing 400044, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peiyi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biorheological Science and Technology, Ministry of Education and Bioengineering College, Chongqing University, Chongqing 400044, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daijing","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biorheological Science and Technology, Ministry of Education and Bioengineering College, Chongqing University, Chongqing 400044, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qiong","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Basic Medicine, Chongqing Medical and Pharmaceutical College, Chongqing 401331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6877-8677","authenticated-orcid":false,"given":"Jun","family":"Yang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biorheological Science and Technology, Ministry of Education and Bioengineering College, Chongqing University, Chongqing 400044, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1038\/s41378-025-01098-9","article-title":"Integrated 3D microstructured digital microfluidic platform for advanced 3D cell culture","volume":"11","author":"Chen","year":"2025","journal-title":"Microsyst. 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