{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T11:56:33Z","timestamp":1765454193449,"version":"3.46.0"},"reference-count":22,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T00:00:00Z","timestamp":1765411200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Medical Sciences"],"abstract":"<jats:p>Background. Routine lymphocyte counting requires quality assurance validation using quality controls (QCs) that closely resemble fresh human blood leukocytes. This study aimed to evaluate a novel artificial product designed to mimic leukocyte light scatters and marker expression. Methods. FlowCytes and TruCytes, \u201cartificial cell mimics\u201d (Slingshot Biosciences, Emeryville, CA, USA), were tested on CE-IVD-certified systems, namely FACSCanto, FACSLyric (BD Biosciences), Navios, and DxFlex (Beckman Coulter), using routine staining, lysing, fixation, and no-wash procedures for T, B, and NK counting. Results. FlowCytes and TruCytes provided forward and side scatter profiles comparable to human leukocytes on the FACSCanto, FACSLyric, and DxFlex systems but not on Navios despite adapting the process to be slightly different from the manufacturer\u2019s recommendations. TruCytes demonstrated robust immunolabeling of CD3, CD4, CD8, and CD19 on the FACSCanto, FACSLyric, and DxFlex systems, with fluorescence intensities and subset distributions being similar to those usually observed in fresh human blood. However, CD16 and CD56 labeling was inconsistent and depended on the antibody clones used. Regrettably, monocyte and granulocyte mimics lacked expression of CD4, CD16, and CD14. TruCytes also displayed significantly lower concentrations of TBNK lymphocyte subsets compared to healthy human blood. Conclusions. FlowCytes and TruCytes show promises as internal quality controls for T cell and B cell immunophenotyping, but not NK cells. They are compatible with most CE-IVD cytometers, even when using lysis\/fixation\/no-wash routine diagnosis procedures. Further multicentric studies are warranted to assess their performance relative to existing products, such as stabilized human blood.<\/jats:p>","DOI":"10.3390\/medsci13040314","type":"journal-article","created":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T11:17:27Z","timestamp":1765451847000},"page":"314","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of Cell Mimics as Potential Quality Controls for Human Leukocyte Immunophenotyping"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4941-2415","authenticated-orcid":false,"given":"Louis","family":"Waeckel","sequence":"first","affiliation":[{"name":"Laboratoire d\u2019Immunologie, CHU Saint-Etienne, 42055 Saint-Etienne Cedex 2, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brigitte","family":"Le Mauff","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie, CHU Caen, 14000 Caen, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacques","family":"Trauet","sequence":"additional","affiliation":[{"name":"Institut d\u2019Immunologie Laboratoire de D\u00e9ficits Immunitaires et Immunoth\u00e9rapies, CHRU de Lille, F-59000 Lille, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6960-4521","authenticated-orcid":false,"given":"Julie","family":"Demaret","sequence":"additional","affiliation":[{"name":"Institut d\u2019Immunologie Laboratoire de D\u00e9ficits Immunitaires et Immunoth\u00e9rapies, CHRU de Lille, F-59000 Lille, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahmed","family":"Boumediene","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie et d\u2019Immunog\u00e9n\u00e9tique, CHU Limoges, 87042 Limoges, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Margarita","family":"Hurtado-Nedelec","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019H\u00e9matologie et Immunologie, APHP, H\u00f4pital Bichat, 75018 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arnaud","family":"Ciree","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie, CHRU Bretonneau Tours, 37044 Tours, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0076-2895","authenticated-orcid":false,"given":"Gwladys","family":"Bourdenet","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie, CHU Amiens Picardie, 80054 Amiens, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claude","family":"Lambert","sequence":"additional","affiliation":[{"name":"Laboratoire d\u2019Immunologie, CHU Saint-Etienne, 42055 Saint-Etienne Cedex 2, France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,11]]},"reference":[{"key":"ref_1","first-page":"38","article-title":"Toward a culture shift in laboratory quality: Application of the full ISO 15189 standard","volume":"47","author":"Farmer","year":"2015","journal-title":"MLO Med. Lab. Obs."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1002\/cyto.b.21079","article-title":"Accreditation of flow cytometry in Europe","volume":"84","author":"Sack","year":"2013","journal-title":"Cytom. Part B Clin. Cytom."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1002\/cyto.990110402","article-title":"Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry","volume":"11","author":"Loken","year":"1990","journal-title":"Cytometry"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1002\/(SICI)1097-0320(19960315)26:1<16::AID-CYTO3>3.0.CO;2-E","article-title":"Use of CD45 fluorescence and side-scatter characteristics for gating lymphocytes when using the whole blood lysis procedure and flow cytometry","volume":"26","author":"Nicholson","year":"1996","journal-title":"Cytometry"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1002\/cyto.b.20005","article-title":"CD3 bright lymphocyte population reveal gammadelta T cells","volume":"61","author":"Lambert","year":"2004","journal-title":"Cytom. Part B Clin. Cytom."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1086\/644509","article-title":"Common features of gammadelta T cells and CD8+ alphabeta T cells responding to human cytomegalovirus infection in kidney transplant recipients","volume":"200","author":"Couzi","year":"2009","journal-title":"J. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e39871","DOI":"10.1097\/MD.0000000000039871","article-title":"Causal relationship between lymphocyte subsets and the risk of sepsis: A Mendelian randomization study","volume":"103","author":"Chen","year":"2024","journal-title":"Medicine"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1097\/QAD.0000000000003743","article-title":"CD4dim CD8bright T cells are inversely associated with neuro-inflammatory markers among people with HIV","volume":"38","author":"Albalawi","year":"2024","journal-title":"AIDS"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1632","DOI":"10.1515\/cclm-2024-1246","article-title":"Flow-cytometric lymphocyte subsets enumeration: Comparison of single\/dual-platform method in clinical laboratory with dual-platform extended PanLeucogating method in reference laboratory","volume":"63","author":"Hu","year":"2025","journal-title":"Clin. Chem. Lab. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3301","DOI":"10.1182\/blood-2011-02-336321","article-title":"Dual-functional capability of CD3+CD56+ CIK cells, a T-cell subset that acquires NK function and retains TCR-mediated specific cytotoxicity","volume":"118","author":"Pievani","year":"2011","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2053","DOI":"10.1002\/eji.1830230902","article-title":"The novel subset of CD14+\/CD16+ blood monocytes exhibits features of tissue macrophages","volume":"23","author":"Fingerle","year":"1993","journal-title":"Eur. J. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Kapellos, T.S., Bonaguro, L., Gemund, I., Reusch, N., Saglam, A., Hinkley, E.R., and Schultze, J.L. (2019). Human Monocyte Subsets and Phenotypes in Major Chronic Inflammatory Diseases. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.02035"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Hurley, A.A. (1997). Quality control in phenotypic analysis by flow cytometry. Curr. Protoc. Cytom., 6.1.1\u20136.1.4.","DOI":"10.1002\/0471142956.cy0601s00"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0022-1759(00)00226-X","article-title":"Validation and quality control of immunophenotyping in clinical flow cytometry","volume":"243","author":"Owens","year":"2000","journal-title":"J. Immunol. Methods"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1016\/j.cll.2007.05.009","article-title":"Quality control in clinical flow cytometry","volume":"27","author":"Oldaker","year":"2007","journal-title":"Clin. Lab. Med."},{"key":"ref_16","first-page":"1","article-title":"Standardization, calibration, and control in flow cytometry","volume":"79","author":"Hoffman","year":"2005","journal-title":"Curr. Protoc. Cytom."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1038\/nrd1659","article-title":"Therapeutic applications of polymeric artificial cells","volume":"4","author":"Chang","year":"2005","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.cyto.2005.07.009","article-title":"Cytometric bead array (CBA) for the measurement of cytokines in urine and plasma of patients undergoing renal rejection","volume":"32","author":"Jimenez","year":"2005","journal-title":"Cytokine"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1111\/j.1432-2277.2004.00048.x","article-title":"Evaluation of flow cytometric panel reactive antibody in renal transplant recipients\u2014Examination of 238 cases of renal transplantation","volume":"18","author":"Ishida","year":"2005","journal-title":"Transpl. Int."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1007\/s00467-015-3145-z","article-title":"Transplant immuno-diagnostics: Crossmatch and antigen detection","volume":"31","author":"South","year":"2016","journal-title":"Pediatr. Nephrol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1002\/cyto.b.21531","article-title":"Laboratory Accuracy Improvement in the UK NEQAS Leucocyte Immunophenotyping Immune Monitoring Program: An Eleven-Year Review via Longitudinal Mixed Effects Modeling","volume":"94","author":"Bainbridge","year":"2018","journal-title":"Cytom. Part B Clin. Cytom."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1002\/cyto.b.21751","article-title":"Acceptable \u201cReal-Life\u201d Variability for Lymphocyte Counts by Flow Cytometry","volume":"96","author":"Ticchioni","year":"2019","journal-title":"Cytom. Part B Clin. Cytom."}],"container-title":["Medical Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3271\/13\/4\/314\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T11:41:15Z","timestamp":1765453275000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3271\/13\/4\/314"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,11]]},"references-count":22,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["medsci13040314"],"URL":"https:\/\/doi.org\/10.3390\/medsci13040314","relation":{},"ISSN":["2076-3271"],"issn-type":[{"value":"2076-3271","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,11]]}}}