Manufacturing Cell Research Articles

A novel mitochondrial pyruvate carrier inhibitor drives stem cell-like memory CAR T cell generation and enhances antitumor efficacy

Adoptive cell transfer with chimeric antigen receptor (CAR) expressing T cells can induce remarkable complete responses in cancer patients. Therapeutic success has been correlated with central and stem cell-like memory T cell subsets in the infusion product which are better able to drive efficient CAR T cell in vivo expansion and long-term persistence. We previously reported that inhibition of the mitochondrial pyruvate carrier (MPC) during mouse CAR T cell culture, induces a memory phenotype and enhances antitumor efficacy against melanoma. Here, we use a novel MPC inhibitor, MITO-66, which robustly induces a stem cell-like memory phenotype in CD19-CAR T cells generated from healthy donors and patients with relapsed/refractory B cell malignancies. MITO-66-conditioned CAR T cells were superior in controlling human pre-B cell acute lymphoblastic leukemia in mice. Following adoptive cell transfer, MITO-66-conditioned CAR T cells maintained a memory phenotype and protected cured mice against tumor rechallenge. Furthermore, in an in vivo B cell leukemia stress model, CD19-CAR T cells generated in the presence of MITO-66 largely outperformed clinical-stage AKT and PI-3Kδ inhibitors. Thus, we provide compelling preclinical evidence that MPC inhibition with MITO-66 during CAR T cell manufacturing dramatically enhances their antitumor efficacy, thereby paving the way to clinical translation.

Optimizing feature extraction and fusion for high-resolution defect detection in solar cells

In this paper, we propose a novel architecture for defect detection in electroluminescent images of polycrystalline silicon solar cells, addressing the challenges posed by subtle and dispersed defects. Our model, based on a modified Swin Transformer, incorporates key innovations that enhance feature extraction and fusion. We replace the conventional self-attention mechanism with a novel group self-attention mechanism, increasing the mAP50:5:95 score from 50.12 % to 52.98 % while reducing inference time from 74 ms to 62 ms. We also introduce a spatial displacement with shift convolution module, replacing the traditional Multi-Layer Perceptron, which further enhances the model's receptive field and improves precision and recall. Additionally, our fast multi-scale feature fusion mechanism effectively combines high-resolution details with high-level semantic features from different network layers, optimizing defect detection accuracy. Experimental results on the PVEL-AD dataset demonstrate that our model achieves the highest mAP50 score of 83.11 % and an F1-Score of 84.33 %, surpassing state-of-the-art models while maintaining a competitive inference time of 66.3 ms. These findings highlight the effectiveness of our innovations in improving defect detection accuracy and computational efficiency, making our model a robust solution for quality assurance in solar cell manufacturing.

Preclinical Evaluation of AZD6422, an Armored Chimeric Antigen Receptor T Cell Targeting CLDN18.2 in Gastric, Pancreatic, and Esophageal Cancer.

CLDN18.2 is a surface membrane protein crucial for maintaining tight junctions in gastric mucosal cells and is highly expressed in gastric, esophageal, and pancreatic cancers. Thus, CLDN18.2 is suited for exploration as a clinical target for chimeric antigen receptor T-cell (CAR-T) therapy in these indications. Although CAR-T therapies show promise, a challenge faced in their development for solid tumors is the immunosuppressive tumor microenvironment, often characterized by the presence of immune and stromal cells secreting high levels of transforming growth factor beta (TGF-β). Addition of TGF-β armoring can potentially expand CAR-T activity in solid tumors. We report on the preclinical development of a CLDN18.2-targeting CAR-T showing effectiveness in CLDN18.2-positive gastric, esophageal, and pancreatic tumor models. The lead lentivirus product contains a unique single-chain variable fragment, CD28 and CD3z costimulatory and signaling domains, and dominant negative TGF-β receptor armoring, enhancing targeting and safety and counteracting suppression. We developed a shortened cell manufacturing process to enhance the potency of the final product, AZD6422. AZD6422 exhibited significant antitumor activity and tolerability in multiple patient-derived tumor xenograft models with various CLDN18.2 and TGF-β levels, as determined by immunohistochemistry. Efficacy of armored CAR-Ts in tumor models with elevated TGF-β was increased in vitro and in vivo. In vitro restimulation assays established greater persistence and cytolytic function of AZD6422 compared with a traditionally manufactured CAR-T. AZD6422 was safe and efficacious in patient-derived, CLDN18.2-positive murine models of gastrointestinal cancers. Our data support further clinical development of AZD6422 for patients with these cancers.

Biomimetic cell stimulation with a graphene oxide antigen-presenting platform for developing T cell-based therapies.

Chimeric antigen receptor (CAR)-engineered T cells represent a front-line therapy for cancers. However, the current CAR T cell manufacturing protocols do not adequately reproduce immunological synapse formation. Here, in response to this limitation, we have developed a flexible graphene oxide antigen-presenting platform (GO-APP) that anchors antibodies onto graphene oxide. By decorating anti-CD3 (αCD3) and anti-CD28 (αCD28) on graphene oxide (GO-APP3/28), we achieved remarkable T cell proliferation. In vitro interactions between GO-APP3/28 and T cells closely mimic the in vivo immunological synapses between antigen-presenting cells and T cells. This immunological synapse mimicry shows a high capacity for stimulating T cell proliferation while preserving their multifunctionality and high potency. Meanwhile, it enhances CAR gene-engineering efficiency, yielding a more than fivefold increase in CAR T cell production compared with the standard protocol. Notably, GO-APP3/28 stimulated appropriate autocrine interleukin-2 (IL-2) in T cells and overcame the in vitro reliance on external IL-2 supplementation, offering an opportunity to culture T cell-based products independent of IL-2 supplementation.

A Study on Operator Allocation in Consideration of Fatigue in Cell Manufacturing System

A Study on Operator Allocation in Consideration of Fatigue in Cell Manufacturing System

Quality Assessment of Perovskite Solar Cells: An Industrial Point of View

The mass production of photovoltaic (PV) devices requires fast and reliable characterization methods and equipment. PV manufacturers produce a complete module roughly every 20 s, and the electrical performance assessment is typically completed in less than 1 s. Times are even more stringent during cell manufacturing. To be competitive in the PV market, perovskite solar cells and modules aim to the same target, i.e., fast and reliable quality assessment. This communication report discusses the limit of characterizing the current perovskite technology. Standard current vs voltage measurements are compared to maximum power point tracking (MPPT), and a fast MPPT procedure is developed to meet the highly demand standard for quality control in the industry of PV production.

3D Printing as a Strategy to Scale-Up Biohybrid Hydrogels for T Cell Manufacture.

The emergence of cellular immunotherapy treatments is introducing more efficient strategies to combat cancer as well as autoimmune and infectious diseases. However, the cellular manufacturing procedures associated with these therapies remain costly and time-consuming, thus limiting their applicability. Recently, lymph-node-inspired PEG-heparin hydrogels have been demonstrated to improve primary human T cell culture at the laboratory scale. To go one step further in their clinical applicability, we assessed their scalability, which was successfully achieved by 3D printing. Thus, we were able to improve primary human T cell infiltration in the biohybrid PEG-heparin hydrogels, as well as increase nutrient, waste, and gas transport, resulting in higher primary human T cell proliferation rates while maintaining the phenotype. Thus, we moved one step further toward meeting the requirements needed to improve the manufacture of the cellular products used in cellular immunotherapies.

3-Methylthiophene: A Sustainable Solvent for High-Performance Organic Solar Cells.

The use of harmful halogenated or aromatic solvents such as chloroform (CF), chlorobenzene (CB), and o-xylene (o-XY) is one of the greatest barriers to the industrial-scale manufacturing of high-performance organic solar cells (OSCs). Therefore, it is necessary to eliminate the effects of these solvents to ensure practical feasibility of OSCs. We found that the anthracene-terminated polymer donor and small-molecule acceptor BO-4Cl had good solubility in 3-methylthiophene (3-MeT). There were no toxicity labels in the SDS and exposure control limits for 3-MeT. An overall power conversion efficiency of 16.87% was achieved by using 3-MeT as the solvent for solar cell fabrication, which was higher than that of the cells made from CF (16.18%) and o-XY (15.69%). The best OSC based on PM6:D18:L8-BO and fabricated with 3-MeT exhibited a high PCE of 18.13%, which is one of the highest values for cells fabricated from halogen-free solvents. These results indicate that 3-MeT is an eco-friendly and low-toxicity solvent for the sustainable fabrication of the OSC active layer.

Microbial metabolite-guided CAR T cell engineering enhances anti-tumor immunity via epigenetic-metabolic crosstalk.

Emerging data have highlighted a correlation between microbiome composition and cancer immunotherapy outcome. While commensal bacteria and their metabolites are known to modulate the host environment, contradictory effects and a lack of mechanistic understanding impede the translation of microbiome-based therapies into the clinic. In this study, we demonstrate that abundance of the commensal metabolite pentanoate is predictive for survival of chimeric antigen receptor (CAR) T cell patients in two independent cohorts. Its implementation in the CAR T cell manufacturing workflow overcomes solid tumor microenvironments in immunocompetent cancer models by hijacking the epigenetic-metabolic crosstalk, reducing exhaustion and promoting naive-like differentiation. While synergy of clinically relevant drugs mimicked the phenotype of pentanoate-engineered CAR T cells in vitro, in vivo challenge showed inferior tumor control. Metabolic tracing of 13C-pentanoate revealed citrate generation in the TCA cycle via the acetyl- and succinyl-CoA entry points as a unique feature of the C5 aliphatic chain. Inhibition of the ATP-citrate lyase, which links metabolic output and histone acetylation, led to accumulation of pentanoate-derived citrate from the succinyl-CoA route and decreased functionality of SCFA-engineered CAR T cells. Our data demonstrate that microbial metabolites are incorporated as epigenetic imprints and implementation into CAR T cell production might serve as embodiment of the microbiome-host axis benefits for clinical applications.

Self-learning and autonomously adapting manufacturing equipment for the circular factory

Abstract The integration of both linear and circular processes in one production system poses significant challenges. In particular, the reprocessing of end-of-life products is associated with uncertainties at all levels of the production system, from the initial planning and control through to the executing production hardware and intralogistics. To address these challenges, this article presents approaches for self-learning and autonomously adapting production equipment for the Circular Factory. Initially, hardware and software solutions are developed to cover the necessary processes. Reprocessing is covered by modular and reconfigurable manufacturing cells, which also include new process chains such as the combination of additive-subtractive processes. The provided capabilities must be applied to ever new products, for example by transferring human procedures for unknown products to the production equipment. Lastly, an overall robust and dynamic production planning and control system is developed that maintains continuous operation even in unforeseen situations. The resulting highly dynamic overall system is connected by an autonomous intralogistics system.

Easy to fabricate 3D metastructure for low-frequency vibration control

As a burgeoning category of elastic metamaterials, 3D metastructures have garnered significant research attention for manipulating low-frequency acoustic and elastic waves. Bandgap engineering allows for the control of these waves across a subwavelength ultrawide frequency range. However, the manufacturing of these 3D structures poses a challenge, necessitating additional support materials for 3D-printed components, creating difficulties in mass production. In this study, we propose a novel lightweight 3D metastructure design that is easy to fabricate and provides a low-frequency subwavelength bandgap. We replaced conventional struts supporting heavy mass inclusions in typical designs with modified arch beams. This structural modification enables the easy and self-supporting manufacturing of 3D metastructure unit cells without the need for extra support material. Utilizing magnets and steel masses with bolts as hard inclusions, the magnet facilitates the quick assembly of the 3D metastructure, potentially facilitating mass manufacturing in practical applications. The wave dispersion and bandgap properties of the metastructure are investigated numerically, and experimental vibration tests are performed on the 3D-printed and assembled parts. The experimental results and numerical findings demonstrate robust vibration attenuation at low frequencies by the proposed 3D metastructure. The suggested, easy-to-fabricate 3D-metastructure design holds potential applications in low-frequency elastic-wave manipulation, including noise and vibration control.

Heterogeneous Nucleation and Enhanced Charge Transfer via Amorphous Metal‐Organic Frameworks for Efficient and Stable Perovskite Solar Cells

AbstractThe two‐step sequential deposition method is considered a potential way for the large‐scale manufacture of perovskite solar cells (PSCs) with high power conversion efficiency and reproducibility. However, the dense lead iodide (PbI2) film interferes with its full contact with organic solutions, resulting in an inadequate reaction at the interface. Herein, 2 kinds of metal‐organic framework (MOF) are introduced, amorphous Ni‐MOF‐74 (amNi‐MOF‐74) and crystalline Zn‐MOF‐74 (crZn‐MOF‐74), into PbI2 for regulating crystallization. Compared to crZn‐MOF‐74, the incorporation of amNi‐MOF‐74 exhibited rapid nucleation, resulting in high‐quality perovskite films with large grain size, low trap density, and enhanced charge transfer between the perovskite and charge transfer layers. Meanwhile, the content of unstable phase PbI2 left in perovskite films due to insufficient reaction is also reduced. The amNi‐MOF‐74 modified PSCs exhibited a champion power conversion efficiency of 24.17% with good humidity and thermal stability. The unencapsulated device maintains 90% of its initial efficiency after 1000 h storage in dark ambient conditions with ≈30% relative humidity. This strategy provides an effective approach for promoting the crystallization process of perovskite and fabricating efficient and stable PSCs.

Green cell scheduling and performance management of manufacturing system based on heat loss optimization

Heat loss is an important loss in the manufacturing process, which has a significant impact on the overall system performance and resource utilization efficiency. Therefore, optimizing heat loss can not only improve manufacturing efficiency, but also promote the green transformation of enterprises. The purpose of this paper is to explore the green unit scheduling and performance management strategy of manufacturing system based on heat loss optimization, so as to improve the energy efficiency and overall performance of manufacturing system. A new scheduling model is proposed, which combines heat loss analysis with green management strategy. By using multi-objective optimization algorithm, the scheduling of manufacturing cells is optimized, focusing on the minimization of heat loss and the maximization of production efficiency. The thermal energy loss characteristics of each process and the operating state of the equipment are considered in the model, and the energy efficiency performance of different scheduling schemes is verified by simulation experiments. The experimental results show that the optimized scheduling scheme significantly reduces the thermal energy loss of the manufacturing system, significantly improves the overall production efficiency, and effectively improves the green performance indicators, including the reduction of energy consumption and waste emission. Through the scheduling and management strategy based on the optimization of heat loss, the green production goal of the manufacturing system is realized. The research of this paper provides a new idea and method for manufacturing enterprises to realize sustainable development while pursuing economic benefit.

Temperature impact on acoustic wave reflection in quasi-collinear acousto-optic devices.

Quasi-collinear geometry is a special configuration of acousto-optic (AO) diffraction that applies the acoustic wave reflection from the AO cell input optical face and provides an extremely large interaction length for achieving abnormally high spectral resolution of AO tunable filters. As a result, it becomes possible to implement the multifrequency diffraction which has found important applications for laser pulse shaping. The operation of quasi-collinear AO devices in the multifrequency diffraction regimen is accompanied by the appearance of the longitudinal and transverse temperature gradients in the crystal, mainly due to the acoustic power absorption. Temperature changes the AO cell material stiffness moduli, affecting the characteristics of the incident and reflected acoustic waves (propagation velocities and walk-off angles), and the reflection condition in general. On the example of paratellurite crystal is shown that the AO cell heating near the reflecting facet leads to a deviation of the reflected acoustic beam propagation direction from that specified during the AO cell manufacturing. The deviation magnitude depends on the reflection geometry choice and, in the paratellurite, may exceed several degrees, which adversely affects the AO diffraction characteristics, reducing the AO interaction efficiency and distorting the transmission function shape. The reflected beam deviation may be compensated by means of choosing the angle between the AO cell reflecting face and the piezoelectric transducer face, taking into account the operating AO device thermal regimen.

Immunosuppressant therapy averts rejection of allogeneic FKBP1A-disrupted CAR-T cells

Chimeric antigen receptor (CAR) Tcells from allogeneic donors promise "off-the-shelf" availability by overcoming challenges associated with autologous cell manufacturing. However, recipient immunologic rejection of allogeneic CAR-T cells may decrease their invivo lifespan and limit treatment efficacy. Here, we demonstrate that the immunosuppressants rapamycin and tacrolimus effectively mitigate allorejection of HLA-mismatched CAR-T cells in immunocompetent humanized mice, extending their invivo persistence to that of syngeneic humanized mouse-derived CAR-T cells. In turn, genetic knockout (KO) of FKBP prolyl isomerase 1A (FKBP1A), which encodes a protein targeted by both drugs, was necessary to confer CD19-specific CAR-T cells (19CAR) robust functional resistance to these immunosuppressants. FKBP1AKO 19CAR-T cells maintained potent invitro functional profiles and controlled invivo tumor progression similarly to untreated 19CAR-T cells. Moreover, immunosuppressant treatment averted invivo allorejection permitting FKBP1AKO 19CAR-T cell-driven B cell aplasia. Thus, we demonstrate that genome engineering enables immunosuppressant treatment to improve the therapeutic potential of universal donor-derived CAR-T cells.

Unity and ROS as a Digital and Communication Layer for Digital Twin Application: Case Study of Robotic Arm in a Smart Manufacturing Cell.

A digital twin (DT) is a virtual/digital model of any physical object (physical twin), interconnected through data exchange. In the context of Industry 4.0, DTs are integral to intelligent automation driving innovation at scale by providing significant improvements in precision, flexibility, and real-time responsiveness. A critical challenge in developing DTs is achieving a model that reflects real-time conditions with precision and flexibility. This paper focuses on evaluating latency and accuracy, key metrics for assessing the efficacy of a DT, which often hinder scalability and adaptability in robotic applications. This article presents a comprehensive framework for developing DTs using Unity and Robot Operating System (ROS) as the main layers of digitalization and communication. The MoveIt package was used for motion planning and execution for the robotic arm, showcasing the framework's versatility independent of proprietary constraints. Leveraging the versatility and open-source nature of these tools, the framework ensures interoperability, adaptability, and scalability, crucial for modern smart manufacturing applications. Our approach was validated by conducting extensive accuracy and latency tests. We measured latency by timestamping messages exchanged between the physical and digital twin, achieving a latency of 77.67 ms. Accuracy was assessed by comparing the joint positions of the DT and the physical robotic arm over multiple cycles, resulting in an accuracy rate of 99.99%. The results highlight the potential of DTs in enhancing operational efficiency and decision-making in manufacturing environments.

Biofunctionalization of Cellulose Microcarriers Using a Carbohydrate Binding Module Linked with Fibroblast Growth Factor for the Expansion of Human Umbilical Mesenchymal Stromal Cells in Stirred Suspension Bioreactors.

Mesenchymal stromal cells (MSCs) have the potential to be used as autologous or allogenic cell therapy in several diseases due to their beneficial secretome and capacity for immunomodulation and differentiation. However, clinical trials using MSCs require a large number of cells. As an alternative to traditional culture flasks, suspension bioreactors provide a scalable platform to produce clinically relevant quantities of cells. When cultured in bioreactors, anchorage-dependent cells like MSCs require the addition of microcarriers, which provide a surface for cell attachment while in suspension. The best performing microcarriers are typically coated in animal derived proteins, which increases cellular attachment and proliferation but present issues from a regulatory perspective. To overcome this issue, a recombinant fusion protein was generated linking basic fibroblast growth factor (bFGF) to a cellulose-specific carbohydrate binding module (CBM) and used to functionalize the surface of cellulose microcarriers for the expansion of human umbilical MSCs in suspension bioreactors. The fusion protein was shown to support the growth of MSCs when used as a soluble growth factor in the absence of cellulose, readily bound to cellulose microcarriers in a dose-dependent manner, and ultimately improved the expansion of MSCs when grown in bioreactors using cellulose microcarriers. The use of CBM fusion proteins offers a simple method for the surface immobilization of growth factors to animal component-free substrates such as cellulose, which can be used alongside bioreactors to increase growth factor lifespan, decrease culture medium cost, and increase cell production in the manufacturing of therapeutic cells.

How photovoltaic industry policies foster the development of silicon solar cell manufacturing technology: Based on Self-attention mechanism

With the advancement of silicon solar cell manufacturing technology (SSCM-Tec) driven by subsidy policies, some developing countries have implemented subsidy reduction policies. Concurrently, intense international competition has prompted the implementation of restriction policies. However, due to SSCM involving multiple manufacturing steps, each step comprising various SSCM-Tec, it presents challenges in researching how different types of policies affect SSCM-Tec in each step. We construct the SSCM-Mechanism based on patent claims to capture the nuanced changes in SSCM-Tec caused by the implementation of policies. The results show that: (1) Policies lead to an imbalance in SSCM-Tec advancements among manufacturing steps; (2) Different types of policies have varying impacts on SSCM-Tec. Supportive policies boost enterprises' interest in developing SSCM-Tec, and restrictive policies and subsidy reduction policies speed up SSCM-Tec innovation; (3) The collaboration of various policy types can better drive the advancement of SSCM-Tec. Employing the right type of policy at the opportune moment serves as a crucial driving force for fostering innovation in SSCM-Tec. These findings reveal the multifaceted impact of policies on SSCM-Tec, providing policymakers with a clearer perspective to formulate more precise policies for guiding the direction of SSCM-Tec development and accelerating its innovation pace.

Fluorine substitution for aggregation transformation and exciton dissociation acceleration in non-fused electron acceptors

In order to enable low-cost manufacture of organic solar cells (OSCs), it is very important to develop non-fused electron acceptors to match with one of the cheapest polymer donors, poly(3-hexylthiophene) (P3HT). However, the lower exciton dissociation and charge transfer are still the main factors that limit the improvement of P3HT-based OSCs. Herein, we designed one class of A2-A1-D-A1-A2 type non-fused electron acceptors and synthesized BTA34 and F-BTA34 by introducing benzo[d][1,2,3] triazole as the bridged unit. The dominated H-aggregation in F-BTA34 affords high degree of crystalline and proper phase separation when blended with polymer P3HT, which ensure efficient exciton dissociation and higher charge carrier mobility. Consequently, P3HT:F-BTA34 devices demonstrate a better performance with higher short-circuit current and fill factor. The blend of P3HT:BTA34 exhibits a broader distribution of the mixing domains, resulting in relative longer-lived charge species. This work provides useful strategies to design non-fused electron acceptors to match with P3HT from the perspective of simultaneously improving exciton dissociation and suppressing charge recombination.

CD19 CAR T cells for B cell malignancies: a systematic review and meta-analysis focused on clinical impacts of CAR structural domains, manufacturing conditions, cellular product, doses, patient’s age, and tumor types

CD19-targeted chimeric antigen receptors (CAR) T cells are one of the most remarkable cellular therapies for managing B cell malignancies. However, long-term disease-free survival is still a challenge to overcome. Here, we evaluated the influence of different hinge, transmembrane (TM), and costimulatory CAR domains, as well as manufacturing conditions, cellular product type, doses, patient’s age, and tumor types on the clinical outcomes of patients with B cell cancers treated with CD19 CAR T cells. The primary outcome was defined as the best complete response (BCR), and the secondary outcomes were the best objective response (BOR) and 12-month overall survival (OS). The covariates considered were the type of hinge, TM, and costimulatory domains in the CAR, CAR T cell manufacturing conditions, cell population transduced with the CAR, the number of CAR T cell infusions, amount of CAR T cells injected/Kg, CD19 CAR type (name), tumor type, and age. Fifty-six studies (3493 patients) were included in the systematic review and 46 (3421 patients) in the meta-analysis. The overall BCR rate was 56%, with 60% OS and 75% BOR. Younger patients displayed remarkably higher BCR prevalence without differences in OS. The presence of CD28 in the CAR’s hinge, TM, and costimulatory domains improved all outcomes evaluated. Doses from one to 4.9 million cells/kg resulted in better clinical outcomes. Our data also suggest that regardless of whether patients have had high objective responses, they might have survival benefits from CD19 CAR T therapy. This meta-analysis is a critical hypothesis-generating instrument, capturing effects in the CD19 CAR T cells literature lacking randomized clinical trials and large observational studies.