Cytotherapy最新文献

{"title":"INTER-DONOR VARIABILITY AND CELLULAR PROLIFERATION: THEIR IMPACT ON THE IMMUNOMODULATORY STRENGTH OF ADIPOSE-DERIVED MESENCHYMAL STEM CELLS IN T CELL-MEDIATED HEPATITIS","authors":"C.X. Chan ,&nbsp;T. Okubo ,&nbsp;M. Haga ,&nbsp;H. Kurata ,&nbsp;S. Ota ,&nbsp;Y. Ueno ,&nbsp;H. Kawai ,&nbsp;M. Hashimoto ,&nbsp;Y. Honma","doi":"10.1016/j.jcyt.2024.03.064","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.03.064","url":null,"abstract":"<div><h3>Background &amp; Aim</h3><p>Mesenchymal stem cells (MSCs) are recognized for their numerous therapeutic benefits, notably for their immunomodulatory capabilities. Thus, they are being tested in many clinical trials, including those for GVHD and osteoarthritis. However, MSCs are also known to be highly heterogeneous. In addition to originating from various tissue sources, they also exhibit inter-donor variation within the same tissue, making it hard to standardise MSC therapeutics. Hence, our aim is to improve this by investigating donor variability and the link to their immunomodulatory strength.</p></div><div><h3>Methods, Results &amp; Conclusion</h3><p>We tested this using both in-house produced and commercially sold adipose-derived MSCs (AD-MSCs) via the Concanavalin A hepatitis animal model. In both sets of AD-MSCs, we indeed observed varying levels of anti-inflammatory effects across different donors. Microarray analysis on our in-house AD-MSCs revealed donors with stronger effects exhibited activation in cell cycling and proliferation pathways. Single cell transcriptomic analysis on the commercially sold AD-MSCs, however, showed all donors merging into one group regardless of effect strength, indicating a high level of similarity in MSC characteristics. Further examination through cell clustering revealed 12 distinct subclusters that made up different percentages within each donor, suggesting that subtle cell population differences play a crucial role in differentiating AD-MSCs with strong and weak anti-inflammatory effects. Gene ontology analysis on the differentially expressed genes (DEGS) from clusters representing donors with strong effects demonstrated changes in cell proliferation, echoing the results of the in-house AD-MSC microarray analysis. When we assessed the in vitro doubling time (DT) and cell growth in both sets of AD-MSCs, we observed that donors with stronger anti-inflammatory effects indeed exhibited lower DT and higher growth rates. In summary, this study is the first to link the in vivo immunomodulatory effects of different donors to bulk and single cell transcriptomics, with results indicating a positive correlation between cell proliferation and effect strength. Our ongoing research includes further dissecting target subclusters within each donor and identifying common DEGs between both in-house and commercial AD-MSCs that could serve as reliable Critical Quality Attribute (CQA) markers. This will hopefully lead to better standardisation and development of more effective AD-MSC therapeutics.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141078270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"IL-27 IN A FOURTH GENERATION CAR ENHANCES CAR-NK CELL EFFECTOR FUNCTION","authors":"A.F. Biggi ,&nbsp;R.N. Silvestre ,&nbsp;M.C. Tirapelle ,&nbsp;J.T. de Azevedo ,&nbsp;D. Covas ,&nbsp;K.C. Malmegrim ,&nbsp;M.L. Figueiredo ,&nbsp;V. Picanço-Castro","doi":"10.1016/j.jcyt.2024.03.015","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.03.015","url":null,"abstract":"<div><h3>Background &amp; Aim</h3><p>Interleukin 27 (IL-27), part of the IL-12 cytokine family, enhances NK cell cytotoxicity by inducing activating receptors (e.g., NKp46, NKG2D), and stimulating IFN-gamma production. While chimeric antigen receptor (CAR) has shown promise for NK cells in early-phase clinical trials, improving CAR-NK cell therapeutic effectiveness is critical. Although transgenic expression of cytokines (e.g., IL-15) in CAR-NK cells partially enhances pro-survival or proliferative properties, further research is needed to identify the optimal cytokine. Therefore, investigating the role of co-expressing IL-27 with a fourth-generation CD19-targeted CAR on NK cells is crucial.</p></div><div><h3>Methods, Results &amp; Conclusion</h3><p>Lentiviral particles carrying CAR19 and CAR19-IL27 vectors were used to transduce NK-92 cell line. CAR+ cells were FACS-sorted and the potency of CAR-NK cells was evaluated against CD19+ cell lines (NALM-6 and Raji) both <em>in vitro</em> and in <em>in vivo</em> murine models. Tumor burden was quantified through bioluminescence. RNA from target-activated CAR19 and CAR19-IL27 was prepared following Illumina's TruSeq-stranded-mRNA protocol and conducted by LC Sciences. Our findings suggest that CAR19-IL27 enhances NK-92 inherent proliferative ability by about 4-fold compared to CAR19. Furthermore, CAR19-IL27 cells exhibited higher <em>in vitro</em> cytotoxicity against NALM-6 and Raji compared to CAR19, p&lt;0.005 (NALM-6) and p&lt;0.00005 (Raji); higher activation-induced degranulation (CD107a MFI), and IFN-gamma secretion; and decreased expression of inhibitory checkpoint CTLA-4 at both protein (-20-fold) and RNA levels (-1,45 log2FC). Potentially, IL-27 enhances NK cell cytotoxicity through MAPK-related pathways, as genes associated with this pathway were upregulated in CAR19-IL27 cells (e.g., <em>CCR7, ICAM-1, IGF2, NTRK2</em>, and <em>NTRK3</em>). Furthermore, <em>in vivo</em> experiments show that CAR19-IL27 NK cells, in contrast to CAR19 cells, exhibit an extended ability to control tumor growth, reducing tumor burden on day 21 (4 mice/group). Therefore, integrating IL-27 into CAR-NK cells is a promising strategy to boost therapeutic responses against cancer cells, supporting the evidence of fourth-generation CAR engineering in advancing NK cell-based immunotherapies. Financially supported by FAPESP 2020/07055-9, 2014/50947-7, 2013/08135-2; CNPq 440543/2022-3; CAPES 88887.817043/2023-00.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141078282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FEASIBILITY, SAFETY, AND EFFICACY OF AUTOLOGOUS ADIPOSE-DERIVED MESENCHYMAL STROMAL CELLS ADHERED TO TYPE I/III COLLAGEN MEMBRANE FOR FOCAL KNEE CARTILAGE DEFECTS: A PHASE 1 CLINICAL TRIAL","authors":"C.C. Kaleka ,&nbsp;E. Antonioli ,&nbsp;N.C. Martins-Oliveira ,&nbsp;P.D. Silva ,&nbsp;A.D. Castro ,&nbsp;M. Cohen","doi":"10.1016/j.jcyt.2024.03.058","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.03.058","url":null,"abstract":"<div><h3>Background &amp; Aim</h3><p>Mesenchymal stromal cell (MSC) transplantation has shown promising outcomes in treating focal chondral defects of the knee, providing notable improvements in pain and functional scores. This study aims to assess the feasibility, safety, and efficacy of a Tissue Engineering Product (TEP) utilizing autologous adipose tissue (AT)-MSCs associated to a type I/III collagen membrane for the treatment of symptomatic focal cartilage defects in knee.</p></div><div><h3>Methods, Results &amp; Conclusion</h3><p>Four patients with full-thickness knee cartilage defects received TEP implantation and were prospectively followed by 12 months. Autologous subcutaneous adipose tissue was harvested and transported to a private cell culture facility (StemCorp), where it was processed for cell isolation and expansion under a GMP system. Once characterized, the AT-MSC cells were associated to collagen type I/III membrane. Quality control tests were performed along the manufacture and for final TEP release. Knee surgery for TEP implantation was performed by arthrotomy, with the chondral defect prepared and the TEP sutured over the lesion with the help of fibrin glue. After surgery, all patients followed the same rehabilitation protocol. Safety of the procedure was assessed by the incidence of reoperations for any cause after TEP implantation, and by incidence and severity of complications. Efficacy was evaluated by means of clinical scales for functional status: International Knee Documentation Committee (IKDC); Knee injury and Osteoarthritis Outcome Score (KOOS); Lysholm Knee Scoring Scale, SF36 for quality life and Visual Analogue Scale (VAS) for pain intensity. Besides, patients were evaluated by magnetic resonance imaging (RMI) preoperatively and 12 months postoperatively. The study demonstrated the safety and feasibility of the TEP, with no reoperations or joint stiffness reported. After 12 months, substantial improvements were observed in functional scales (IKDC: 138%, KOOS: 92%, Lysholm: 53%), pain reduction (-72%), and quality of life (SF36 social functioning domain: 23%). Post-surgical MRI confirmed satisfactory tissue filling, resembling hyaline cartilage around the lesion according to the MOCART (Magnetic Resonance Observation of Cartilage Repair Tissue) scale. This Phase 1 trial suggests that autologous AT-MSCs on a collagen membrane present a safe and effective approach for treating knee cartilage defects. The promising results warrant further exploration in larger clinical studies.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141077852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HIGHLY SPECIFICITY OF EFFECTOR CELLS TO INDUCE DIFFERENT TYPE OF NEURAL PROGENITOR CELLS FROM AMSC","authors":"N. Mahajam ,&nbsp;K. Gordon ,&nbsp;M. Ladd ,&nbsp;M. Schwartz ,&nbsp;J. Vaughan ,&nbsp;G.A. Moviglia","doi":"10.1016/j.jcyt.2024.03.098","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.03.098","url":null,"abstract":"<div><h3>Background &amp; Aim</h3><p>In 2006 it was published that lymphocytes, with autoimmune effector activity (EC), against the Central Nervous System (CNS) induce MSCs to differentiate into neural progenitor cells. Furthermore, both cell types combined were able to repair damage to the CNS. Subsequently, a similar phenomenon was found in the coincubation of EC against cartilage, bone, skeletal muscle, retina and recently in smooth muscle and urothelium on aMSC. In each case, MSCs became progenitor cells of the tissue against which ECs were activated. To study the degree of specificity of ECs with respect to their induction of cellular and tissue differentiation, the following experimental plan was proposed.</p></div><div><h3>Methods, Results &amp; Conclusion: Materials and methods</h3><p>3 similar but different antigens (of porcine origin) were identified: whole brain extract, substantia nigra and autonomic splanchnic ganglion. Next, they challenged MNC from human blood (obtained through volunteer red cross donors) to obtain EC against these 3 different tissues. MNCs were incubated in bottles without attachment at concentration of 3.2 million per ml, in DMEM high glucose, enriched with 1% tissue antigen (Protein in micrograms). TC medium was enriched with famotidine (4.5 micrograms per ml) and indomethacin (10 micrograms per ml) to inhibit the growth of regulatory clones. Resulting ECs were cocultured with aMSCs of human origin (donated from lipoaspiration surpluses of aesthetic purposes) and placental stem cells (voluntarily donated by disease-free pregnant women at term). MSC differentiation was studied using tissue-specific differentiation markers.</p></div><div><h3>Results</h3><p>After three days, the three cocultures showed cells positive for tubulin 3 (universal marker of young neurons) and for GFAP (universal marker of Glia cells). Only cocultures where was used EC against Substantia Nigra showed positive cells for Tyrosine Hydroxylase (exclusive marker for Dopaminergic cells), and only cocultures that used EC for autonomic cells showed cells with marker TUJ1 and p75, markers for neurons and glia of the autonomic system.</p></div><div><h3>Conclusion</h3><p>The high specificity of ECs to induce the differentiation of various types of neurons leads us to think about a sophisticated form of epigenetic induction that accompanies the tissue-specific activation of effector cells.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141078350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"BEYOND CAR-T: HARNESSING NATURAL KILLER (NK) CELLS FOR IMPROVED OUTCOMES IN HIGH-RISK NEUROBLASTOMA","authors":"T. Baker, K. Schultz","doi":"10.1016/j.jcyt.2024.04.046","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.04.046","url":null,"abstract":"","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141151039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MICROPOROUS PARTICLE SCAFFOLDS FOR MESENCHYMAL STROMAL CELL EXPANSION AND ENHANCED IMMUNOMODULATORY FUNCTION","authors":"T. Spoerer, C. Williams, R. Marklein","doi":"10.1016/j.jcyt.2024.04.020","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.04.020","url":null,"abstract":"","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141151172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"COMPARISON STUDY OF DIFFERENT PUMP TECHNOLOGIES’ IMPACT ON CELL VIABILITY","authors":"N. Mahasena, O. Akintewe, M. Jaroszeski, B. Biller, A. Biller, P. Priebe","doi":"10.1016/j.jcyt.2024.04.039","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.04.039","url":null,"abstract":"","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141153637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ALLOGENEIC IPSC-DERIVED NK THERAPEUTIC PRODUCT: PHENOTYPIC ANALYSIS AND FUNCTIONAL CHARACTERIZATION","authors":"M. Houppermans, L. Lam, M.F. Mata, S. Luharia, A. Habich-Crayton, G. D'Agostino, J. Hasan, T. Ponomaryov, Y. Choo, M. Tarunina","doi":"10.1016/j.jcyt.2024.04.056","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.04.056","url":null,"abstract":"","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141151187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"AGING OF MESENCHYMAL STEM CELLS IN VITRO: IMPLICATIONS IN PHENOTYPE AND DIFFERENTIATION CAPACITY","authors":"M. Sielski ,&nbsp;H. Andrade ,&nbsp;R.D. Alves Paiva ,&nbsp;L. Coa ,&nbsp;D. Oliveira ,&nbsp;J.M. Kutner ,&nbsp;A.T. kondo ,&nbsp;L.N. Kerbauy ,&nbsp;J. Azevedo ,&nbsp;O.K. Okamoto ,&nbsp;J.A. Godoy","doi":"10.1016/j.jcyt.2024.03.078","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.03.078","url":null,"abstract":"<div><h3>Background &amp; Aim</h3><p>Human mesenchymal stem cells (hMSC) have been used in various clinical protocols; these cells are multipotent and can be isolated from various sources. They are able to adhere to plastic, differentiate into mesodermal lineage and express specific markers (CD90⁺/CD73⁺/CD105⁺/CD45^-/CD34^-/CD19^-/CD11b^-/HLA-DR^-). Despite these features and the easily culture method, hMSC present a limited growing time that can affect its use in clinical protocols. In order to stablish the upper limit by which hMSC could be cultivated, hMSC proliferating rate was analyzed from P0 to P16; hMSC were also analyzed regarding its capacity to differentiate into mesodermal cells, imunophenotyping profile, capacity to decrease mononuclear cells proliferation <em>in vitro</em> and its telomere lenght. hMSC was isolated from bone marrow from healthy donors.</p></div><div><h3>Methods, Results &amp; Conclusion</h3><p>Population doubling time was observed during 16 passages; the cumulative doubling time increased during the first 5 passages and seems to be maintained from P5 to P8. After P8, proliferation rate slowed down. Differentiation capacity, imunophenotyping, MNC proliferation assay and telomere lenght were analyzed in P5, P7, P10 and P14 or P15. Differentiation capacity was decreased by P10 when was observed a less differentiation into adipocytes (very few cells were observed with lipid droplets in the cytoplasm), chondrocytes (a less presence of proteolycans) and osteoblasts (less calcium deposits). Related to the markers expression., its results corroborate the ones observed in differentiation assay where there was a decrease in CD73 (55%) and CD105 (73%) expression on P10; CD90 presented a expression higher than 85% in all passages analyzed. The negative markers such as CD19, HLA-DR and CD34 presented a higher expression on P10 (7,53%, 7,73% and 3,11%, respectivelly). The telomere lenght showed a decrease on P10 when compared to the other passages analyzed (8.9 versus 9.5 on P0, 9.4 on P3, 9.6 on P5 and 9.5 on P7). The immunomodulatory analysis also showed a decrease in modulate the MNCs proliferation on passage 10; when analyzing 1 MSC:1 MNC, the number of proliferating cells increased from 12% (P5), 16% (P7) to 57% (P10) and 66% (P15). All these data suggest that hMSC is changing its proliferative into a senescent profile when cells reach the P10. In conclusion, hMSC could be used for clinical purposes until passage 7-8, since passage 10 already showed great modificiations related to potency, imunophenotyping, proliferation and telomere length.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141077733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A FED-BATCH CHEMICALLY DEFINED HMSC-EV BIOPROCESS MEDIUM ENABLING 2-4X EV YIELD IMPROVEMENTS IN BIOREACTOR CULTURE","authors":"T.M. Willstaedt ,&nbsp;A. Walde ,&nbsp;J.A. Rowley","doi":"10.1016/j.jcyt.2024.03.105","DOIUrl":"https://doi.org/10.1016/j.jcyt.2024.03.105","url":null,"abstract":"<div><h3>Background &amp; Aim</h3><p>Extracellular vesicles (EVs) derived from human mesenchymal stromal cells (hMSC-EVs) have been studied in over 200 preclinical applications and dozens of human clinical trials, underscoring the need for scalable production processes compatible with GMP environments. Most existing 2D and 3D Bioreactor hMSC-EV production processes require a cell expansion stage utilizing undefined components, followed by a wash and medium exchange to remove expansion medium impurities prior to an EV collection phase in a defined medium. Simplifying this 3D process to include cell expansion and EV collection in one medium requires chemically defined growth conditions, a fed-batch medium design, and an efficient process to maximize cell and EV yield, and final product quality. We have developed a chemically defined, scalable fed-batch bioreactor production medium to enable the streamlined and highly efficient production of hMSC-EVs. This study evaluates hMSC-EV production and EV quality across multiple donors and tissues in microcarrier spinner flask cultures using a traditional cell expansion, wash, collect process vs the single-step production process, including scale-up to a 3L stirred tank bioreactor.</p></div><div><h3>Methods, Results &amp; Conclusion</h3><p>MSC-EVs were produced from hMSCs (hBM and hUC RoosterVial, 1M) in either RoosterNourish-MSC-XF/RoosterReplenish/RoosterCollect-EV or the new highly productive, chemically defined (HiDef-EV) fed-batch system and collected EVs at set times. HiDef-EV cultures led to increased EV production on days 5, 7, 10 and 12 of culture, while maintaining healthy viable cell profiles. The fed-batch process for hMSC-EV production increased the EV collection window from healthy hMSCs resulting in 2-4x increase in hMSC-EV yield over traditional EV production processes. Elimination of the medium exchange and wash steps resulted in utilization of fewer raw materials, retention rather than disposal of EVs produced during cell growth, and significant reductions in total media used and total cost per billion EVs. Additionally, EV Quality Attributes including size, tetraspanin expression, CD-73 activity, RNA, and lipid content are preserved in the HiDef-EV system. Scale up in 3L Eppendorf bioreactor showed comparable cell growth, EV yields and EV quality between traditional and HiDef-EV process. This highly productive chemically defined EV medium is a simplified, time and cost saving solution for the large-scale production of higher purity hMSC-EVs necessary for extensive clinical investigations.</p></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141078275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}