{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,25]],"date-time":"2026-02-25T22:39:38Z","timestamp":1772059178154,"version":"3.50.1"},"reference-count":53,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2021,4,22]],"date-time":"2021-04-22T00:00:00Z","timestamp":1619049600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Macrophages are innate immune cells that play critical roles in tissue homeostasis, inflammation, and immune oncology. Macrophages differentiated from human induced pluripotent stem cells (iPSCs) overcome many limitations of using peripheral blood derived macrophages. The ability to scale up and cryopreserve a large amount of end stage macrophages from single clonal iPSCs from normal and disease specific donors offers a unique opportunity for genomic analysis and drug screening. The present study describes the step wise generation and characterization of macrophages from iPSCs using a defined serum free method amenable to scale up to generate a large batch of pure end stage cryopreservable macrophages expressing CD68, CD33, CD11c, CD11b, CD1a, HLA-DR, CD86, CD64, CD80, CD206, CD169, CD47, HLA-ABC, and CX3CR. The end stage macrophages pre and post cryopreservation retain purity, morphology, responsiveness to stimuli and display robust phagocytic function coming right out of cryopreservation. The same differentiation process was used to generate end stage macrophages from isogenic iPSCs engineered to mimic mutations associated with Parkinson\u2019s disease (SNCA A53T), neuronal ceroid lipofuscinosis (GRN2\/GRN R493X), and Rett syndrome (MECP2-Knockout). End stage macrophages from isogenic engineered clones displayed differential macrophage-specific purity markers, phagocytic function, and response to specific stimuli. Thus, generating a panel of functional, physiologically relevant iPSC-derived macrophages can potentially facilitate the understanding of neural inflammatory responses associated with neurodegeneration.<\/jats:p>","DOI":"10.1371\/journal.pone.0250107","type":"journal-article","created":{"date-parts":[[2021,4,22]],"date-time":"2021-04-22T18:36:09Z","timestamp":1619116569000},"page":"e0250107","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":9,"title":["Generation of cryopreserved macrophages from normal and genetically engineered human pluripotent stem cells for disease modelling"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1060-7143","authenticated-orcid":true,"given":"Christie","family":"Munn","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah","family":"Burton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah","family":"Dickerson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kiranmayee","family":"Bakshy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anne","family":"Strouse","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8820-4328","authenticated-orcid":true,"given":"Deepika","family":"Rajesh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,4,22]]},"reference":[{"issue":"3\u20134","key":"pone.0250107.ref001","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1007\/s00424-017-1945-7","article-title":"Physiological roles of macrophages","volume":"469","author":"S Gordon","year":"2017","journal-title":"Pflugers Arch"},{"issue":"3","key":"pone.0250107.ref002","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.ymeth.2007.06.004","article-title":"Macrophage scavenger receptors and host-derived ligands","volume":"43","author":"A Pluddemann","year":"2007","journal-title":"Methods"},{"issue":"6","key":"pone.0250107.ref003","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1038\/nri3671","article-title":"Monocytes and macrophages: developmental pathways and tissue homeostasis","volume":"14","author":"F Ginhoux","year":"2014","journal-title":"Nat Rev Immunol"},{"issue":"1","key":"pone.0250107.ref004","doi-asserted-by":"crossref","first-page":"2552","DOI":"10.1038\/s41467-020-16355-w","article-title":"Peripheral nerve resident macrophages share tissue-specific programming and features of activated microglia","volume":"11","author":"PL Wang","year":"2020","journal-title":"Nat Commun"},{"issue":"12","key":"pone.0250107.ref005","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1038\/nri1733","article-title":"Monocyte and macrophage heterogeneity","volume":"5","author":"S Gordon","year":"2005","journal-title":"Nat Rev Immunol"},{"issue":"1","key":"pone.0250107.ref006","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.immuni.2012.12.001","article-title":"Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis","volume":"38","author":"S Yona","year":"2013","journal-title":"Immunity"},{"issue":"3","key":"pone.0250107.ref007","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1007\/s10571-019-00651-1","article-title":"Apolipoprotein E Polymorphism and Oxidative Stress in Peripheral Blood-Derived Macrophage-Mediated Amyloid-Beta Phagocytosis in Alzheimer\u2019s Disease Patients","volume":"39","author":"PS Jairani","year":"2019","journal-title":"Cell Mol Neurobiol"},{"issue":"18","key":"pone.0250107.ref008","doi-asserted-by":"crossref","DOI":"10.3390\/ijms20184559","article-title":"Niacin Ameliorates Neuro-Inflammation in Parkinson\u2019s Disease via GPR109A","volume":"20","author":"B Giri","year":"2019","journal-title":"Int J Mol Sci"},{"key":"pone.0250107.ref009","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.jneuroim.2018.05.002","article-title":"Niacin modulates macrophage polarization in Parkinson\u2019s disease","volume":"320","author":"C Wakade","year":"2018","journal-title":"J Neuroimmunol"},{"issue":"1","key":"pone.0250107.ref010","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1038\/nrn3406","article-title":"The many faces of alpha-synuclein: from structure and toxicity to therapeutic target","volume":"14","author":"HA Lashuel","year":"2013","journal-title":"Nat Rev Neurosci"},{"issue":"2","key":"pone.0250107.ref011","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1007\/s12035-012-8369-3","article-title":"The central theme of Parkinson\u2019s disease: alpha-synuclein","volume":"47","author":"M Ozansoy","year":"2013","journal-title":"Mol Neurobiol"},{"issue":"Suppl 1","key":"pone.0250107.ref012","doi-asserted-by":"crossref","first-page":"S16","DOI":"10.1016\/j.parkreldis.2015.08.015","article-title":"Phenotypic spectrum of alpha-synuclein mutations: New insights from patients and cellular models","volume":"22","author":"S Petrucci","year":"2016","journal-title":"Parkinsonism Relat Disord"},{"key":"pone.0250107.ref013","doi-asserted-by":"crossref","first-page":"65","DOI":"10.4103\/sni.sni_441_16","article-title":"Parkinson\u2019s disease: Microglial\/macrophage-induced immunoexcitotoxicity as a central mechanism of neurodegeneration","volume":"8","author":"RL Blaylock","year":"2017","journal-title":"Surg Neurol Int"},{"issue":"1","key":"pone.0250107.ref014","doi-asserted-by":"crossref","first-page":"9003","DOI":"10.1038\/s41598-017-09362-3","article-title":"Excess alpha-synuclein compromises phagocytosis in iPSC-derived macrophages","volume":"7","author":"W Haenseler","year":"2017","journal-title":"Sci Rep"},{"issue":"11","key":"pone.0250107.ref015","doi-asserted-by":"crossref","first-page":"1245","DOI":"10.1002\/bies.200900086","article-title":"The granulin gene family: from cancer to dementia","volume":"31","author":"A Bateman","year":"2009","journal-title":"Bioessays"},{"issue":"1","key":"pone.0250107.ref016","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1093\/brain\/awz377","article-title":"Homozygous GRN mutations: new phenotypes and new insights into pathological and molecular mechanisms","volume":"143","author":"V Huin","year":"2020","journal-title":"Brain"},{"issue":"9","key":"pone.0250107.ref017","doi-asserted-by":"crossref","first-page":"2611","DOI":"10.1084\/jem.20160999","article-title":"Progranulin deficiency causes impairment of autophagy and TDP-43 accumulation","volume":"214","author":"MC Chang","year":"2017","journal-title":"J Exp Med"},{"issue":"1","key":"pone.0250107.ref018","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.gene.2012.12.009","article-title":"Regulation of the MIR155 host gene in physiological and pathological processes","volume":"532","author":"TS Elton","year":"2013","journal-title":"Gene"},{"issue":"3","key":"pone.0250107.ref019","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1016\/j.jpeds.2005.10.037","article-title":"Rett syndrome in Australia: a review of the epidemiology","volume":"148","author":"CL Laurvick","year":"2006","journal-title":"J Pediatr"},{"issue":"10","key":"pone.0250107.ref020","doi-asserted-by":"crossref","first-page":"3377","DOI":"10.1007\/s10803-015-2519-1","article-title":"Brief report: systematic review of Rett syndrome in males","volume":"45","author":"B Reichow","year":"2015","journal-title":"J Autism Dev Disord"},{"issue":"4","key":"pone.0250107.ref021","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.immuni.2015.04.002","article-title":"Brains, Blood, and Guts: MeCP2 Regulates Microglia, Monocytes, and Peripheral Macrophages","volume":"42","author":"DP Schafer","year":"2015","journal-title":"Immunity"},{"issue":"1","key":"pone.0250107.ref022","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/S1090-3798(02)00134-4","article-title":"Neurodevelopmental disorders in males related to the gene causing Rett syndrome in females (MECP2)","volume":"7","author":"U Moog","year":"2003","journal-title":"Eur J Paediatr Neurol"},{"issue":"5858","key":"pone.0250107.ref023","doi-asserted-by":"crossref","first-page":"1917","DOI":"10.1126\/science.1151526","article-title":"Induced pluripotent stem cell lines derived from human somatic cells","volume":"318","author":"J Yu","year":"2007","journal-title":"Science"},{"issue":"15","key":"pone.0250107.ref024","doi-asserted-by":"crossref","first-page":"1987","DOI":"10.1101\/gad.1689808","article-title":"Pluripotent stem cell lines","volume":"22","author":"J Yu","year":"2008","journal-title":"Genes Dev"},{"issue":"11","key":"pone.0250107.ref025","doi-asserted-by":"crossref","first-page":"e27956","DOI":"10.1371\/journal.pone.0027956","article-title":"Generation of induced pluripotent stem cells from CD34+ cells across blood drawn from multiple donors with non-integrating episomal vectors","volume":"6","author":"AA Mack","year":"2011","journal-title":"PLoS One"},{"issue":"9","key":"pone.0250107.ref026","doi-asserted-by":"crossref","first-page":"2818","DOI":"10.1172\/JCI38591","article-title":"Generation of mature human myelomonocytic cells through expansion and differentiation of pluripotent stem cell-derived lin-CD34+CD43+CD45+ progenitors","volume":"119","author":"KD Choi","year":"2009","journal-title":"J Clin Invest"},{"issue":"3","key":"pone.0250107.ref027","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1038\/nprot.2010.184","article-title":"Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells","volume":"6","author":"KD Choi","year":"2011","journal-title":"Nat Protoc"},{"issue":"8","key":"pone.0250107.ref028","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1038\/s41587-019-0201-4","article-title":"Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype","volume":"37","author":"D Kim","year":"2019","journal-title":"Nat Biotechnol"},{"issue":"5","key":"pone.0250107.ref029","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: an R package for comparing biological themes among gene clusters","volume":"16","author":"G Yu","year":"2012","journal-title":"OMICS"},{"key":"pone.0250107.ref030","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1186\/1471-2105-12-152","article-title":"ZFN-site searches genomes for zinc finger nuclease target sites and off-target sites","volume":"12","author":"TJ Cradick","year":"2011","journal-title":"BMC Bioinformatics"},{"issue":"11","key":"pone.0250107.ref031","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1038\/nri3073","article-title":"Protective and pathogenic functions of macrophage subsets","volume":"11","author":"PJ Murray","year":"2011","journal-title":"Nat Rev Immunol"},{"key":"pone.0250107.ref032","doi-asserted-by":"crossref","first-page":"816460","DOI":"10.1155\/2015\/816460","article-title":"Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms","volume":"2015","author":"T Roszer","year":"2015","journal-title":"Mediators Inflamm"},{"issue":"2","key":"pone.0250107.ref033","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1093\/jb\/mvy050","article-title":"CD169 macrophages regulate immune responses toward particulate materials in the circulating fluid","volume":"164","author":"K Asano","year":"2018","journal-title":"J Biochem"},{"issue":"2","key":"pone.0250107.ref034","first-page":"80","article-title":"Macrophage checkpoint blockade: results from initial clinical trials, binding analyses, and CD47-SIRPalpha structure-function","volume":"3","author":"AR Jalil","year":"2020","journal-title":"Antib Ther"},{"issue":"12","key":"pone.0250107.ref035","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1016\/j.it.2017.07.009","article-title":"The Multifaceted Personality of Intestinal CX3CR1(+) Macrophages","volume":"38","author":"M Regoli","year":"2017","journal-title":"Trends Immunol"},{"issue":"1","key":"pone.0250107.ref036","doi-asserted-by":"crossref","first-page":"e5","DOI":"10.4110\/in.2018.18.e5","article-title":"Tissue-specific Role of CX3CR1 Expressing Immune Cells and Their Relationships with Human Disease","volume":"18","author":"M Lee","year":"2018","journal-title":"Immune Netw"},{"key":"pone.0250107.ref037","doi-asserted-by":"crossref","first-page":"1593","DOI":"10.3389\/fimmu.2018.01593","article-title":"CD38 Is Robustly Induced in Human Macrophages and Monocytes in Inflammatory Conditions","volume":"9","author":"SA Amici","year":"2018","journal-title":"Front Immunol"},{"issue":"2","key":"pone.0250107.ref038","article-title":"iPSC-Derived Human Microglia-like Cells to Study Neurological Diseases","volume":"94","author":"EM Abud","year":"2017","journal-title":"Neuron"},{"key":"pone.0250107.ref039","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.bcp.2016.07.011","article-title":"A novel real time imaging platform to quantify macrophage phagocytosis","volume":"116","author":"TS Kapellos","year":"2016","journal-title":"Biochem Pharmacol"},{"issue":"2","key":"pone.0250107.ref040","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.cryobiol.2006.12.005","article-title":"Characterization of rat and human Kupffer cells after cryopreservation","volume":"54","author":"P Walbrun","year":"2007","journal-title":"Cryobiology"},{"key":"pone.0250107.ref041","doi-asserted-by":"crossref","first-page":"2952","DOI":"10.3389\/fimmu.2018.02952","article-title":"Innate Anti-microbial and Anti-chemotaxis Properties of Progranulin in an Acute Otitis Media Mouse Model","volume":"9","author":"Z Wang","year":"2018","journal-title":"Front Immunol"},{"issue":"4","key":"pone.0250107.ref042","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1016\/j.immuni.2015.03.013","article-title":"Methyl-CpG Binding Protein 2 Regulates Microglia and Macrophage Gene Expression in Response to Inflammatory Stimuli","volume":"42","author":"JC Cronk","year":"2015","journal-title":"Immunity"},{"key":"pone.0250107.ref043","doi-asserted-by":"crossref","first-page":"2265","DOI":"10.3389\/fimmu.2019.02265","article-title":"Metabolic Programming of Macrophages: Implications in the Pathogenesis of Granulomatous Disease","volume":"10","author":"JL Wilson","year":"2019","journal-title":"Front Immunol"},{"issue":"12","key":"pone.0250107.ref044","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1016\/j.it.2004.09.015","article-title":"The chemokine system in diverse forms of macrophage activation and polarization","volume":"25","author":"A Mantovani","year":"2004","journal-title":"Trends Immunol"},{"issue":"6","key":"pone.0250107.ref045","doi-asserted-by":"crossref","DOI":"10.3390\/ijms19061801","article-title":"Role of Human Macrophage Polarization in Inflammation during Infectious Diseases","volume":"19","author":"C Atri","year":"2018","journal-title":"Int J Mol Sci"},{"issue":"1","key":"pone.0250107.ref046","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.cellimm.2013.01.010","article-title":"Polarization profiles of human M-CSF-generated macrophages and comparison of M1-markers in classically activated macrophages from GM-CSF and M-CSF origin","volume":"281","author":"M Jaguin","year":"2013","journal-title":"Cell Immunol"},{"issue":"9","key":"pone.0250107.ref047","doi-asserted-by":"crossref","first-page":"6425","DOI":"10.1002\/jcp.26429","article-title":"Macrophage plasticity, polarization, and function in health and disease","volume":"233","author":"A Shapouri-Moghaddam","year":"2018","journal-title":"J Cell Physiol"},{"issue":"1","key":"pone.0250107.ref048","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1186\/s12974-020-01748-x","article-title":"Astroglial TLR9 antagonism promotes chemotaxis and alternative activation of macrophages via modulation of astrocyte-derived signals: implications for spinal cord injury","volume":"17","author":"L Li","year":"2020","journal-title":"J Neuroinflammation"},{"issue":"1","key":"pone.0250107.ref049","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1189\/jlb.1A0314-170R","article-title":"The chemotaxis of M1 and M2 macrophages is regulated by different chemokines","volume":"97","author":"W Xuan","year":"2015","journal-title":"J Leukoc Biol"},{"key":"pone.0250107.ref050","doi-asserted-by":"crossref","first-page":"1930","DOI":"10.3389\/fimmu.2018.01930","article-title":"Chemokine-Induced Macrophage Polarization in Inflammatory Conditions","volume":"9","author":"P Ruytinx","year":"2018","journal-title":"Front Immunol"},{"issue":"7","key":"pone.0250107.ref051","doi-asserted-by":"crossref","first-page":"1619","DOI":"10.1073\/pnas.1621356114","article-title":"Screen for reactivation of MeCP2 on the inactive X chromosome identifies the BMP\/TGF-beta superfamily as a regulator of XIST expression","volume":"114","author":"S Sripathy","year":"2017","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"2","key":"pone.0250107.ref052","doi-asserted-by":"crossref","DOI":"10.3390\/brainsci6020018","article-title":"Neuroimmunology of the Interleukins 13 and 4","volume":"6","author":"S Mori","year":"2016","journal-title":"Brain Sci"},{"issue":"4","key":"pone.0250107.ref053","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1523\/JNEUROSCI.1523-19.2019","article-title":"Convergence between Microglia and Peripheral Macrophages Phenotype during Development and Neuroinflammation","volume":"40","author":"F Grassivaro","year":"2020","journal-title":"J Neurosci"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0250107","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,4,22]],"date-time":"2021-04-22T18:37:01Z","timestamp":1619116621000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0250107"}},"subtitle":[],"editor":[{"given":"Marcel M.","family":"Daadi","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2021,4,22]]},"references-count":53,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4,22]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0250107","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,22]]}}}