Pub Date : 2025-11-14DOI: 10.1016/j.jcyt.2025.101999
Dilpreet Rayat , Jack Miller , Stephanie Richardson-Solorzano , Rylee E. King , Valerie C. West , Alvin W. Su , Justin Parreno
Autologous chondrocyte implantation (ACI) is a cell-based therapy used to treat focal articular cartilage defects. In ACI, chondrocytes from nonload-bearing healthy cartilage regions are isolated and sent to a cell manufacturing laboratory, where they are expanded for cell number in monolayer culture. The expanded cells are then transported back to the clinic for reimplantation into the defect site. The storage and transport conditions from the cell manufacturing facility to the clinic implantation may influence reparative potential. However, the impact of acute hypothermic storage of passaged chondrocyte, which is thought to preserve cell viability, is not fully known. Here, we tested the hypothesis that acute hypothermic storage negatively impacts passaged chondrocyte viability and reduces the capacity for redifferentiation. Passaged chondrocytes were stored either in monolayer culture or in suspension at 36°C, 19°C or 8°C. In monolayer culture, hypothermic temperatures preserved cell viability, promoted cell rounding, reduced proliferation, depolymerized filamentous actin and reduced the mRNA levels of specific matrix molecules compared to 36°C. The effects of hypothermia were context-dependent. Exposure of passaged cells in suspension to hypothermia promoted cell viability and reduced cell aggregation. At 8°C, cells in suspension enhanced expression of specific matrix molecule mRNA levels. Subsequently, when cells in suspension at 8°C were seeded in three-dimensional within agarose molds, aggrecan expression was enhanced and formed thicker tissues. Therefore, in contrast to our hypothesis, we found that hypothermic storage did not have a negative impact; when stored for 1 day in suspension, it had lasting effects on matrix deposition. The storage of passaged chondrocytes under hypothermic conditions may be beneficial for ACI, warranting further investigations of cell hypothermic storage for in vivo repair.
{"title":"The effects of hypothermic storage on passaged chondrocyte viability and redifferentiation potential","authors":"Dilpreet Rayat , Jack Miller , Stephanie Richardson-Solorzano , Rylee E. King , Valerie C. West , Alvin W. Su , Justin Parreno","doi":"10.1016/j.jcyt.2025.101999","DOIUrl":"10.1016/j.jcyt.2025.101999","url":null,"abstract":"<div><div>Autologous chondrocyte implantation (ACI) is a cell-based therapy used to treat focal articular cartilage defects. In ACI, chondrocytes from nonload-bearing healthy cartilage regions are isolated and sent to a cell manufacturing laboratory, where they are expanded for cell number in monolayer culture. The expanded cells are then transported back to the clinic for reimplantation into the defect site. The storage and transport conditions from the cell manufacturing facility to the clinic implantation may influence reparative potential. However, the impact of acute hypothermic storage of passaged chondrocyte, which is thought to preserve cell viability, is not fully known. Here, we tested the hypothesis that acute hypothermic storage negatively impacts passaged chondrocyte viability and reduces the capacity for redifferentiation. Passaged chondrocytes were stored either in monolayer culture or in suspension at 36°C, 19°C or 8°C. In monolayer culture, hypothermic temperatures preserved cell viability, promoted cell rounding, reduced proliferation, depolymerized filamentous actin and reduced the mRNA levels of specific matrix molecules compared to 36°C. The effects of hypothermia were context-dependent. Exposure of passaged cells in suspension to hypothermia promoted cell viability and reduced cell aggregation. At 8°C, cells in suspension enhanced expression of specific matrix molecule mRNA levels. Subsequently, when cells in suspension at 8°C were seeded in three-dimensional within agarose molds, aggrecan expression was enhanced and formed thicker tissues. Therefore, in contrast to our hypothesis, we found that hypothermic storage did not have a negative impact; when stored for 1 day in suspension, it had lasting effects on matrix deposition. The storage of passaged chondrocytes under hypothermic conditions may be beneficial for ACI, warranting further investigations of cell hypothermic storage for <em>in vivo</em> repair.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 2","pages":"Article 101999"},"PeriodicalIF":3.2,"publicationDate":"2025-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145685594","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}
Pub Date : 2025-11-12DOI: 10.1016/j.jcyt.2025.102000
Edwin M. Horwitz
{"title":"Bone marrow grafts on demand","authors":"Edwin M. Horwitz","doi":"10.1016/j.jcyt.2025.102000","DOIUrl":"10.1016/j.jcyt.2025.102000","url":null,"abstract":"","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 2","pages":"Article 102000"},"PeriodicalIF":3.2,"publicationDate":"2025-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145656188","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}
Pub Date : 2025-11-12DOI: 10.1016/j.jcyt.2025.102002
Soo Jung Lee , Gi June Min , Sung-Soo Park , Youngwoo Jeon , Jae Won Yoo , Dong Wook Jekarl , Jae Wook Lee , Ki-Seong Eom , Nack-Gyun Chung , Chang-Ki Min , Seok-Goo Cho , Yonggoo Kim , Jae-Ho Yoon
Background
Tisagenlecleucel (tisa-cel), a CD19-targeted chimeric antigen receptor T-cell (CAR-T) therapy, is an approved treatment for relapsed or refractory (R/R) diffuse large B-cell lymphoma (DLBCL) and B-cell precursor acute lymphoblastic leukemia (B-ALL). While pivotal trials demonstrated its efficacy, real-world evidence from Asian populations remains scarce. In particular, data on manufacturing feasibility, clinical outcomes, and predictive biomarkers in Korean patients are limited.
Objective
This study aimed to evaluate the clinical outcomes and prognostic factors of tisa-cel therapy in Korean patients with R/R DLBCL and B-ALL, with a specific focus on apheresis product quality and baseline immune markers.
Study design
A single-center retrospective study was conducted including 91 patients (DLBCL = 72; B-ALL = 19) intended to receive tisa-cel between April 2022 and April 2024. Apheresis data, treatment responses, survival outcomes, and toxicity profiles were analyzed. The intention-to-treat (ITT) cohort was defined from the time of apheresis. Cox regression was used to identify predictors of survival and relapse, and separate analyses were performed for DLBCL and B-ALL.
Results
Out of 91 patients, 72 received tisa-cel infusion. One-year overall survival (OS) and disease-free survival (DFS) for the entire ITT cohort were 45.9% and 37.5%, respectively. In 19 B-ALL after tisa-cel infusion, 1-year OS and DFS were 68.4% and 52.6%, with no non-relapse mortality (NRM). In 53 DLBCL after tisa-cel infusion, 1-year OS and DFS were 46.8% and 41.6%, respectively, with 29.6% cumulative incidence of relapse among responders. NRM was 9.5% mostly observed in elderly patients. In the DLBCL subgroup, poor OS was independently associated with third-or-later-line salvage therapy (hazard ratio [HR] 2.39), high peripheral blood monocyte proportion at apheresis (>10%, HR 2.57), low CD3+ T-cell concentration in apheresis product (<1.0×10⁸/mL, HR 4.31), and occurrence of immune effector cell-associated neurotoxicity syndrome (HR 2.47). B-ALL patients demonstrated favorable survival and lower toxicity, but no significant prognostic markers were identified.
Conclusions
Tisa-cel therapy was feasible and effective in Korean patients with R/R B-cell malignancies. Pre-treatment immune parameters such as monocyte burden and CD3+ T-cell quality in apheresis products significantly influenced outcomes in DLBCL. These findings suggest that optimizing patient selection and refining manufacturing strategies may enhance CAR-T efficacy, particularly in resource-constrained or heterogeneous real-world settings.
{"title":"Prognostic factors and outcomes of tisagenlecleucel in relapsed or refractory B-cell malignancies: a single-center real-world study","authors":"Soo Jung Lee , Gi June Min , Sung-Soo Park , Youngwoo Jeon , Jae Won Yoo , Dong Wook Jekarl , Jae Wook Lee , Ki-Seong Eom , Nack-Gyun Chung , Chang-Ki Min , Seok-Goo Cho , Yonggoo Kim , Jae-Ho Yoon","doi":"10.1016/j.jcyt.2025.102002","DOIUrl":"10.1016/j.jcyt.2025.102002","url":null,"abstract":"<div><h3>Background</h3><div>Tisagenlecleucel (tisa-cel), a CD19-targeted chimeric antigen receptor T-cell (CAR-T) therapy, is an approved treatment for relapsed or refractory (R/R) diffuse large B-cell lymphoma (DLBCL) and B-cell precursor acute lymphoblastic leukemia (B-ALL). While pivotal trials demonstrated its efficacy, real-world evidence from Asian populations remains scarce. In particular, data on manufacturing feasibility, clinical outcomes, and predictive biomarkers in Korean patients are limited.</div></div><div><h3>Objective</h3><div>This study aimed to evaluate the clinical outcomes and prognostic factors of tisa-cel therapy in Korean patients with R/R DLBCL and B-ALL, with a specific focus on apheresis product quality and baseline immune markers.</div></div><div><h3>Study design</h3><div>A single-center retrospective study was conducted including 91 patients (DLBCL = 72; B-ALL = 19) intended to receive tisa-cel between April 2022 and April 2024. Apheresis data, treatment responses, survival outcomes, and toxicity profiles were analyzed. The intention-to-treat (ITT) cohort was defined from the time of apheresis. Cox regression was used to identify predictors of survival and relapse, and separate analyses were performed for DLBCL and B-ALL.</div></div><div><h3>Results</h3><div>Out of 91 patients, 72 received tisa-cel infusion. One-year overall survival (OS) and disease-free survival (DFS) for the entire ITT cohort were 45.9% and 37.5%, respectively. In 19 B-ALL after tisa-cel infusion, 1-year OS and DFS were 68.4% and 52.6%, with no non-relapse mortality (NRM). In 53 DLBCL after tisa-cel infusion, 1-year OS and DFS were 46.8% and 41.6%, respectively, with 29.6% cumulative incidence of relapse among responders. NRM was 9.5% mostly observed in elderly patients. In the DLBCL subgroup, poor OS was independently associated with third-or-later-line salvage therapy (hazard ratio [HR] 2.39), high peripheral blood monocyte proportion at apheresis (>10%, HR 2.57), low CD3+ T-cell concentration in apheresis product (<1.0×10⁸/mL, HR 4.31), and occurrence of immune effector cell-associated neurotoxicity syndrome (HR 2.47). B-ALL patients demonstrated favorable survival and lower toxicity, but no significant prognostic markers were identified.</div></div><div><h3>Conclusions</h3><div>Tisa-cel therapy was feasible and effective in Korean patients with R/R B-cell malignancies. Pre-treatment immune parameters such as monocyte burden and CD3+ T-cell quality in apheresis products significantly influenced outcomes in DLBCL. These findings suggest that optimizing patient selection and refining manufacturing strategies may enhance CAR-T efficacy, particularly in resource-constrained or heterogeneous real-world settings.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 2","pages":"Article 102002"},"PeriodicalIF":3.2,"publicationDate":"2025-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145652079","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}
CAR-T cell therapy has revolutionized the treatment of non-Hodgkin B-lymphomas, but relapse rates remain unacceptably high, and resistance mechanisms are still unclear. A key challenge of T-cell−based therapy is the immune-suppressive tumor microenvironment, which is rich in immune-suppressive cells, including regulatory T cells (Tregs). Tregs express FOXP3, CD3 and CD25, and their role in B-cell lymphomas treated with CAR-T cells remains uncertain. Using immunohistochemistry, we analyzed 31 pre-infusion and seven post-infusion paired biopsies from patients with relapsed/refractory B-cell lymphoma who received CAR-T cell therapy. Pre- infusion FOXP3+ cell percentages were assessed and correlated with response rate, progression-free survival (PFS), and overall survival (OS). Post-infusion biopsies were examined for FOXP3 changes. We identified 3% as the optimal FOXP3+ cut-off, defining low (≤3%) and high (>3%) FOXP3 groups. Patients with high FOXP3 had a better OS (1-year OS 94% versus 61%, P = 0.006). PFS also favored high FOXP3 patients but lacked statistical significance (1-year PFS 65% versus 54%, P = 0.41). Notably, relapsed patients with high FOXP3 maintained high expression and responded better to subsequent treatments (P = 0.03). Our findings highlight the impact of tumor-infiltrating Tregs on CAR-T cell efficacy in lymphomas.
CAR-T细胞疗法已经彻底改变了非霍奇金b淋巴瘤的治疗，但复发率仍然高得令人无法接受，而且耐药机制仍不清楚。基于T细胞的治疗的一个关键挑战是免疫抑制肿瘤微环境，其中富含免疫抑制细胞，包括调节性T细胞（Tregs）。Tregs表达FOXP3、CD3和CD25，它们在CAR-T细胞治疗的b细胞淋巴瘤中的作用尚不确定。使用免疫组织化学，我们分析了31例输注前和7例输注后的成对活检，这些活检来自接受CAR-T细胞治疗的复发/难治性b细胞淋巴瘤患者。评估输注前FOXP3+细胞百分比，并将其与缓解率、无进展生存期（PFS）和总生存期（OS）相关。注射后活检检查FOXP3变化。我们确定3%为最佳FOXP3+临界值，定义了低（≤3%）和高（低于3%）FOXP3组。FOXP3高的患者有更好的OS（1年OS 94% vs 61%， P = 0.006）。PFS也有利于高FOXP3患者，但缺乏统计学意义（1年PFS 65% vs 54%， P = 0.41）。值得注意的是，FOXP3高表达的复发患者保持高表达，对后续治疗的反应更好（P = 0.03）。我们的研究结果强调了肿瘤浸润性Tregs对CAR-T细胞治疗淋巴瘤疗效的影响。
{"title":"A FOXP3+ cell-rich microenvironment associates with better overall survival after CAR-T cell therapy in relapsed/refractory B-cell lymphoma","authors":"Simone Zanella , Cristina Zucchinetti , Chiara de Philippis , Daniele Mannina , Daniela Taurino , Jacopo Mariotti , Barbara Sarina , Daoud Rahal , Carmelo Carlo-Stella , Armando Santoro , Arturo Bonometti , Stefania Bramanti","doi":"10.1016/j.jcyt.2025.10.007","DOIUrl":"10.1016/j.jcyt.2025.10.007","url":null,"abstract":"<div><div>CAR-T cell therapy has revolutionized the treatment of non-Hodgkin B-lymphomas, but relapse rates remain unacceptably high, and resistance mechanisms are still unclear. A key challenge of T-cell−based therapy is the immune-suppressive tumor microenvironment, which is rich in immune-suppressive cells, including regulatory T cells (Tregs). Tregs express FOXP3, CD3 and CD25, and their role in B-cell lymphomas treated with CAR-T cells remains uncertain. Using immunohistochemistry, we analyzed 31 pre-infusion and seven post-infusion paired biopsies from patients with relapsed/refractory B-cell lymphoma who received CAR-T cell therapy. Pre- infusion FOXP3+ cell percentages were assessed and correlated with response rate, progression-free survival (PFS), and overall survival (OS). Post-infusion biopsies were examined for FOXP3 changes. We identified 3% as the optimal FOXP3+ cut-off, defining low (≤3%) and high (>3%) FOXP3 groups. Patients with high FOXP3 had a better OS (1-year OS 94% versus 61%, <em>P</em> = 0.006). PFS also favored high FOXP3 patients but lacked statistical significance (1-year PFS 65% versus 54%, <em>P</em> = 0.41). Notably, relapsed patients with high FOXP3 maintained high expression and responded better to subsequent treatments (<em>P</em> = 0.03). Our findings highlight the impact of tumor-infiltrating Tregs on CAR-T cell efficacy in lymphomas.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101997"},"PeriodicalIF":3.2,"publicationDate":"2025-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145524579","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}
Pub Date : 2025-11-01DOI: 10.1016/j.jcyt.2025.10.005
Jeremy Snyder
Background Aims
Cellular and gene therapy products provide hope to people experiencing debilitating diseases. Many people seek to access these products ahead of regulatory approval through clinical trials, exceptional access programs, and direct to consumer purchases. Even when regulatory approval has been given, these products can cost millions of dollars in direct and indirect expenses. This study examines how crowdfunding campaigns are being used to pay for United States Federal Drug Administration (FDA) approved cellular and gene therapy products.
Methods
The author searched the GoFundMe crowdfunding platform for 43 FDA-approved cellular and gene therapy products. The campaign title, campaign text, funding requested, funding pledged, number of donations, campaign start date, and campaign location was collected for each campaign. Total 529 campaigns were identified and 322 included for content analysis as seeking funding to pay for the direct or indirect costs of accessing FDA-approved cellular or gene therapy products.
Results
Crowdfunding campaigns for 17 FDA-approved cellular and gene therapy products were identified. Total 322 campaigns raised $8,997,107.19 (median $3675.04) from 154,230 (72.5) donations out of $334,723,452.76 ($184,250) requested. Two campaigns raised 39.2% of all funds and 10 raised 64.8% of all funds. Fundraising goals ranged from $1.11–$12,000,000. The 4th and 5th largest quintiles of fundraising requests were over $1,200,000.
Conclusions
The median fundraising goal and funding received were larger than has been found in other studies of medical crowdfunding. Differences in fundraising goals and donations suggest inequities in insurance coverage and the ability to access cellular and gene therapy products.
{"title":"Crowdfunding for cellular and gene therapy products","authors":"Jeremy Snyder","doi":"10.1016/j.jcyt.2025.10.005","DOIUrl":"10.1016/j.jcyt.2025.10.005","url":null,"abstract":"<div><h3>Background Aims</h3><div>Cellular and gene therapy products provide hope to people experiencing debilitating diseases. Many people seek to access these products ahead of regulatory approval through clinical trials, exceptional access programs, and direct to consumer purchases. Even when regulatory approval has been given, these products can cost millions of dollars in direct and indirect expenses. This study examines how crowdfunding campaigns are being used to pay for United States Federal Drug Administration (FDA) approved cellular and gene therapy products.</div></div><div><h3>Methods</h3><div>The author searched the GoFundMe crowdfunding platform for 43 FDA-approved cellular and gene therapy products. The campaign title, campaign text, funding requested, funding pledged, number of donations, campaign start date, and campaign location was collected for each campaign. Total 529 campaigns were identified and 322 included for content analysis as seeking funding to pay for the direct or indirect costs of accessing FDA-approved cellular or gene therapy products.</div></div><div><h3>Results</h3><div>Crowdfunding campaigns for 17 FDA-approved cellular and gene therapy products were identified. Total 322 campaigns raised $8,997,107.19 (median $3675.04) from 154,230 (72.5) donations out of $334,723,452.76 ($184,250) requested. Two campaigns raised 39.2% of all funds and 10 raised 64.8% of all funds. Fundraising goals ranged from $1.11–$12,000,000. The 4<sup>th</sup> and 5<sup>th</sup> largest quintiles of fundraising requests were over $1,200,000.</div></div><div><h3>Conclusions</h3><div>The median fundraising goal and funding received were larger than has been found in other studies of medical crowdfunding. Differences in fundraising goals and donations suggest inequities in insurance coverage and the ability to access cellular and gene therapy products.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 1","pages":"Article 101995"},"PeriodicalIF":3.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145580047","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}
Pub Date : 2025-10-29DOI: 10.1016/j.jcyt.2025.10.008
Inês Serrenho , Sofia C. Serra , António J. Salgado , Graça Baltazar
Background aims
Neonatal hypoxic-ischemic encephalopathy (HIE) remains a leading cause of infant mortality and long-term neurologic disability. Although therapeutic hypothermia (TH) is the current standard treatment, its effectiveness is limited, with many infants either ineligible or showing incomplete recovery. As a result, alternative therapies, such as mesenchymal stem cells (MSCs) derived from induced pluripotent stem cells (iMSCs), are being explored. iMSCs have shown considerable promise in preclinical studies because of their consistent properties, which help overcome some of the challenges associated with traditional MSCs. This study evaluated the efficacy of iMSCs and their secretome in promoting recovery after neonatal hypoxic-ischemic (HI) brain injury in a well-established rodent model.
Methods
The Rice-Vannucci model was used to induce HI brain injury to the developing brain on postnatal day 10 (P10) Wistar Han rat pups. Two days post-injury, 50 000 iMSCs or their secretome were administered intranasally. Over the next 30 days, motor and cognitive functions were assessed through a series of behavioral tests. In addition, brain lesion size, neurogenesis and glial reactivity were examined by immunohistochemistry to evaluate the extent of neurorepair and inflammation.
Results
HI-lesioned animals treated with intranasal iMSCs or their secretome demonstrated improved motor capabilities and enhanced recognition memory compared with untreated lesioned animals. Both iMSCs and their secretome administration led to a reduction in brain lesion size and increased neurogenesis in the hippocampus. Moreover, glial reactivity, including astrocyte and microglia activation, was significantly reduced 30 days after injury, suggesting a more favorable neuroinflammatory environment in treated groups.
Conclusions
These findings highlight the potential of iMSCs and their secretome to enhance functional recovery and reduce brain damage after neonatal HI. The similar therapeutic outcomes achieved with the iMSC secretome suggest that the secreted factors play a central role in driving neurorepair. More studies are still necessary to gain a better understanding of the mechanisms underlying iMSCs' neuroprotective and neuroreparative effects.
{"title":"Brain on the mend: induced mesenchymal stem cell−based therapies promote functional recovery in rats with neonatal hypoxic-ischemic brain injury","authors":"Inês Serrenho , Sofia C. Serra , António J. Salgado , Graça Baltazar","doi":"10.1016/j.jcyt.2025.10.008","DOIUrl":"10.1016/j.jcyt.2025.10.008","url":null,"abstract":"<div><h3>Background aims</h3><div>Neonatal hypoxic-ischemic encephalopathy (HIE) remains a leading cause of infant mortality and long-term neurologic disability. Although therapeutic hypothermia (TH) is the current standard treatment, its effectiveness is limited, with many infants either ineligible or showing incomplete recovery. As a result, alternative therapies, such as mesenchymal stem cells (MSCs) derived from induced pluripotent stem cells (iMSCs), are being explored. iMSCs have shown considerable promise in preclinical studies because of their consistent properties, which help overcome some of the challenges associated with traditional MSCs. This study evaluated the efficacy of iMSCs and their secretome in promoting recovery after neonatal hypoxic-ischemic (HI) brain injury in a well-established rodent model.</div></div><div><h3>Methods</h3><div>The Rice-Vannucci model was used to induce HI brain injury to the developing brain on postnatal day 10 (P10) Wistar Han rat pups. Two days post-injury, 50 000 iMSCs or their secretome were administered intranasally. Over the next 30 days, motor and cognitive functions were assessed through a series of behavioral tests. In addition, brain lesion size, neurogenesis and glial reactivity were examined by immunohistochemistry to evaluate the extent of neurorepair and inflammation.</div></div><div><h3>Results</h3><div>HI-lesioned animals treated with intranasal iMSCs or their secretome demonstrated improved motor capabilities and enhanced recognition memory compared with untreated lesioned animals. Both iMSCs and their secretome administration led to a reduction in brain lesion size and increased neurogenesis in the hippocampus. Moreover, glial reactivity, including astrocyte and microglia activation, was significantly reduced 30 days after injury, suggesting a more favorable neuroinflammatory environment in treated groups.</div></div><div><h3>Conclusions</h3><div>These findings highlight the potential of iMSCs and their secretome to enhance functional recovery and reduce brain damage after neonatal HI. The similar therapeutic outcomes achieved with the iMSC secretome suggest that the secreted factors play a central role in driving neurorepair. More studies are still necessary to gain a better understanding of the mechanisms underlying iMSCs' neuroprotective and neuroreparative effects.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101998"},"PeriodicalIF":3.2,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145551696","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}
Pub Date : 2025-10-25DOI: 10.1016/j.jcyt.2025.10.006
Jonida Kokiçi , Helena Arellano-Ballestero , Benjamin Hammond , Anucha Preechanukul , Noshin Hussain , Kelly da Costa , Jessica Davies , Sadiyah Mukhtar , Thomas Fernandez , Sabine Kinloch , Fiona M. Burns , Patrick Kennedy , Douglas MacDonald , Mala K. Maini , Upkar S. Gill , Alan Xiaodong Zhuang , Mark W. Lowdell , Karl-Johan Malmberg , Ebba Sohlberg , Dimitra Peppa
Background
Hepatitis B virus (HBV) infection remains a significant global health challenge, leading to chronic liver disease and hepatocellular carcinoma (HCC). Natural killer (NK) cells play an important role in the clearance of HBV-infected cells, but their efficacy is often compromised during chronic infection. Adaptive NK cells, characterized by NKG2C expression and enhanced functional responses, represent a promising therapeutic avenue for enhancing anti-HBV immunity and responses to HBV-driven cancers.
Methods
We applied an established protocol, involving K562-HLA-E expressing feeder cells and cytokines (IL-2), for the expansion of adaptive NK cells from cryopreserved T- and B cell depleted peripheral blood mononuclear cells (PBMCs) derived from donors with chronic HBV infection alone or with Human Immunodeficiency Virus (HIV) co-infection. We evaluated the adaptive profile of expanded NK cells, their antibody-dependent cellular cytotoxicity (ADCC) capacity and functional responses against hepatoma cell lines in the presence or absence of HBV infection.
Results
Expanded NK cells achieved >97% purity, with the NKG2C positive population exhibiting a mean 100-fold expansion. These cells demonstrated a predominantly adaptive phenotype with high surface expression of NKG2C and cytotoxic potential (Granzyme B). They maintained high levels of CD16 surface expression and upregulated CD2, essential for ADCC. Functionally, expanded adaptive NK cells showed enhanced ADCC capacity and functional responses to K562 targets, naive, HBV integrant-expressing, and de novo infected hepatoma cell lines. TGF-β preconditioning induced tissue-resident features (CD103, CD49a) in expanded adaptive NK cells, while preserving their adaptive phenotype and functionality, enhancing their potential for liver targeted immunotherapy. Further, expanded adaptive NK cells demonstrated minimal reactivity against autologous activated T cells, suggesting limited off-target effects.
Conclusions
Our study demonstrates the first successful expansion of adaptive NK cells with robust functional responses from donors with chronic viral infection. This approach creates opportunities for NK cell-based therapies alone or in combination with monoclonal antibodies contributing to HBV functional cure strategies and the treatment of HBV-driven cancers.
{"title":"Expanded adaptive NKG2C+ NK cells exhibit potent ADCC and functional responses against HBV-infected hepatoma cell lines","authors":"Jonida Kokiçi , Helena Arellano-Ballestero , Benjamin Hammond , Anucha Preechanukul , Noshin Hussain , Kelly da Costa , Jessica Davies , Sadiyah Mukhtar , Thomas Fernandez , Sabine Kinloch , Fiona M. Burns , Patrick Kennedy , Douglas MacDonald , Mala K. Maini , Upkar S. Gill , Alan Xiaodong Zhuang , Mark W. Lowdell , Karl-Johan Malmberg , Ebba Sohlberg , Dimitra Peppa","doi":"10.1016/j.jcyt.2025.10.006","DOIUrl":"10.1016/j.jcyt.2025.10.006","url":null,"abstract":"<div><h3>Background</h3><div>Hepatitis B virus (HBV) infection remains a significant global health challenge, leading to chronic liver disease and hepatocellular carcinoma (HCC). Natural killer (NK) cells play an important role in the clearance of HBV-infected cells, but their efficacy is often compromised during chronic infection. Adaptive NK cells, characterized by NKG2C expression and enhanced functional responses, represent a promising therapeutic avenue for enhancing anti-HBV immunity and responses to HBV-driven cancers.</div></div><div><h3>Methods</h3><div>We applied an established protocol, involving K562-HLA-E expressing feeder cells and cytokines (IL-2), for the expansion of adaptive NK cells from cryopreserved T- and B cell depleted peripheral blood mononuclear cells (PBMCs) derived from donors with chronic HBV infection alone or with Human Immunodeficiency Virus (HIV) co-infection. We evaluated the adaptive profile of expanded NK cells, their antibody-dependent cellular cytotoxicity (ADCC) capacity and functional responses against hepatoma cell lines in the presence or absence of HBV infection.</div></div><div><h3>Results</h3><div>Expanded NK cells achieved >97% purity, with the NKG2C positive population exhibiting a mean 100-fold expansion. These cells demonstrated a predominantly adaptive phenotype with high surface expression of NKG2C and cytotoxic potential (Granzyme B). They maintained high levels of CD16 surface expression and upregulated CD2, essential for ADCC. Functionally, expanded adaptive NK cells showed enhanced ADCC capacity and functional responses to K562 targets, naive, HBV integrant-expressing, and <em>de novo</em> infected hepatoma cell lines. TGF-β preconditioning induced tissue-resident features (CD103, CD49a) in expanded adaptive NK cells, while preserving their adaptive phenotype and functionality, enhancing their potential for liver targeted immunotherapy. Further, expanded adaptive NK cells demonstrated minimal reactivity against autologous activated T cells, suggesting limited off-target effects.</div></div><div><h3>Conclusions</h3><div>Our study demonstrates the first successful expansion of adaptive NK cells with robust functional responses from donors with chronic viral infection. This approach creates opportunities for NK cell-based therapies alone or in combination with monoclonal antibodies contributing to HBV functional cure strategies and the treatment of HBV-driven cancers.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 3","pages":"Article 101996"},"PeriodicalIF":3.2,"publicationDate":"2025-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145710268","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}
Natural killer (NK) cell therapies hold great promise for cancer treatment; however, donor-to-donor heterogeneity in the ex vivo expansion process remains a critical bottleneck in their supply. This study aimed to identify factors influencing donor variability in a 2-week-long ex vivo NK cell expansion from peripheral blood mononuclear cells, analyzed across three donors. Single-cell transcriptomics was applied to investigate the distribution of cell types and phenotypes, as well as trajectory inference and differential gene expression. Our results identified that several factors were associated with the variability in the final NK cell fraction and expansion, and that their influence was prevalent between culture days 3 and 8. Compared to a high final NK cell expansion, a culture with a low final NK cell expansion exhibited an upregulation of some stress and inflammatory genes and an increase in one specific subcluster of NK cells already on culture day 3. It showed a low score of CD56Bright CD16– phenotype and a high score of CD56Dim CD16+ phenotype. It also had a decreased presence of cytotoxic CD8+ Tm cells. Among the observed subclusters of CD8+ Tm cells, it exhibited a higher presence of a subcluster associated with a less differentiated and less cytotoxic phenotype, as well as a lower prevalence of a subcluster associated with chemokine and cytotoxic genes. Finally, it had a major expansion of one of the CD8+ Tm cells subclusters annotated as NK-like T cell and characterized by a high CCR5 mRNA expression, while the levels of CCL3, CCL4 and CCL5 mRNA were downregulated. The present findings point toward a potential link between CCL signaling and improved NK cell expansion performance, including possible markers for further investigations, and suggest future strategies to increase the final NK cell fraction and expansion based on donor-specific markers.
{"title":"Single-cell transcriptomics reveals dynamics of natural killer cell expansion in a feeder cell-free culture of peripheral blood mononuclear cells—implications for immunotherapy","authors":"Brian Ladd , Markella Zacharouli , Per-Henrik Holmqvist , Stefanie Renken , Pontus Blomberg , Véronique Chotteau","doi":"10.1016/j.jcyt.2025.10.004","DOIUrl":"10.1016/j.jcyt.2025.10.004","url":null,"abstract":"<div><div>Natural killer (NK) cell therapies hold great promise for cancer treatment; however, donor-to-donor heterogeneity in the <em>ex vivo</em> expansion process remains a critical bottleneck in their supply. This study aimed to identify factors influencing donor variability in a 2-week-long <em>ex vivo</em> NK cell expansion from peripheral blood mononuclear cells, analyzed across three donors. Single-cell transcriptomics was applied to investigate the distribution of cell types and phenotypes, as well as trajectory inference and differential gene expression. Our results identified that several factors were associated with the variability in the final NK cell fraction and expansion, and that their influence was prevalent between culture days 3 and 8. Compared to a high final NK cell expansion, a culture with a low final NK cell expansion exhibited an upregulation of some stress and inflammatory genes and an increase in one specific subcluster of NK cells already on culture day 3. It showed a low score of CD56<sup>Bright</sup> CD16<sup>–</sup> phenotype and a high score of CD56<sup>Dim</sup> CD16<sup>+</sup> phenotype. It also had a decreased presence of cytotoxic CD8<sup>+</sup> Tm cells. Among the observed subclusters of CD8<sup>+</sup> Tm cells, it exhibited a higher presence of a subcluster associated with a less differentiated and less cytotoxic phenotype, as well as a lower prevalence of a subcluster associated with chemokine and cytotoxic genes. Finally, it had a major expansion of one of the CD8<sup>+</sup> Tm cells subclusters annotated as NK-like T cell and characterized by a high CCR5 mRNA expression, while the levels of CCL3, CCL4 and CCL5 mRNA were downregulated. The present findings point toward a potential link between CCL signaling and improved NK cell expansion performance, including possible markers for further investigations, and suggest future strategies to increase the final NK cell fraction and expansion based on donor-specific markers.</div></div>","PeriodicalId":50597,"journal":{"name":"Cytotherapy","volume":"28 1","pages":"Article 101994"},"PeriodicalIF":3.2,"publicationDate":"2025-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145624293","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号