scholarly journals P09.13 Optimization of a GMP-grade large-scale expansion protocol for cytokine-induced killer cells using gas-permeable static culture flasks

2020 ◽  
Vol 8 (Suppl 2) ◽  
pp. A58.2-A59
Author(s):  
A Ventura ◽  
P Palmerini ◽  
A Dalla Pietà ◽  
R Sommaggio ◽  
G Astori ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Cell Density ◽  
Large Scale ◽  
Scale Production ◽  
Static Culture ◽  
Cik Cells ◽  
Standard Culture ◽  
Cik Cell ◽  
Scale Expansion ◽  
Cells Cultured

BackgroundCytokine-Induced Killer (CIK) cells are ex vivo expanded T cells with NK cell phenotype. They express both CD3 and CD56 antigens, and exert a potent antitumor activity against a variety of tumors. Several clinical trials demonstrated the safety and the feasibility of CIK cell therapy, with very low side effects and minimal graft-versus-host toxicity. In this study, we developed a GMP-compliant protocol for robust large-scale expansion of CIK cells using G-Rex® gas-permeable static culture flasks.Materials and MethodsCIK cells were obtained by stimulating healthy donor PBMCs with GMP-grade IFN-γ, IL-2 and CD3 mAbs, and were cultured in G-Rex6® or G-Rex®6M well plates. CIK cells in G-Rex6® were split only once at day 7 to reduce cell density, whereas the number of CIK cells culterd in G-Rex®6M was not adjusted. In both culture conditions, fresh IL-2 was provided every 3–4 days. We compared these two culture protocols with the culture in standard flasks. Phenotype was analyzed by flow cytometry and cytotoxicity was assessed against several tumor cell lines by calcein-release assay.ResultsCIK cells cultured in G-Rex6® well plates showed an outstanding cell expansion compared to G-Rex®6M well plates or standard culture flasks, with a 400-fold expansion and a mean of 109 total cells obtained per single well in 14 days, starting from just 2.5 × 106 cells per well. Moreover, the cultures in G-Rex6® were characterized by an higher percentage of CD3+CD56+ cells, as compared to G-Rex®6M or standard culture flasks. Cells cultured in all devices had a comparable expression of NKG2D, NKp30, NKp44, 2B4 receptors. Importantly, CIK cells expanded in G-Rex®6 were as cytotoxic as cells expanded in standard culture flasks. Conversely, CIK cells cultured in G-Rex®6M showed a remarkable reduction of cytotoxicity against tumor cell targets, thus suggesting that cell density during expansion could affect CIK cell activity.ConclusionsWe propose a GMP-compliant protocol for robust large-scale production of CIK cells. G-Rex® system allows to obtain large amounts of CIK cells highly enriched in the CD3+CD56+ subset and endowed with high cytotoxic activity; this can be accomplished with just a single cell culture split at day 7, which dramatically reduces the culture manipulation as compared to the standard culture flasks. Notably, this strategy can be further and easily scalable to produce CIK cells for clinical immunotherapy applications.Disclosure InformationA. Ventura: None. P. Palmerini: None. A. Dalla Pietà: None. R. Sommaggio: None. G. Astori: None. K. Chieregato: None. M. Tisi: None. C. Visco: None. O. Perbellini: None. M. Ruggeri: None. E. Cappuzzello: None. A. Rosato: None.

Adaptable stirred-tank culture strategies for large scale production of multicellular spheroid-based tumor cell models

2016 ◽  
Vol 221 ◽  
pp. 118-129 ◽  
Author(s):  
Vítor E. Santo ◽  
Marta F. Estrada ◽  
Sofia P. Rebelo ◽  
Sofia Abreu ◽  
Inês Silva ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Large Scale ◽  
Stirred Tank ◽  
Scale Production ◽  
Multicellular Spheroid ◽  
Cell Models ◽  
Tank Culture ◽  
Large Scale Production ◽  
Culture Strategies

scholarly journals Clinical Studies Applying Cytokine-Induced Killer Cells for the Treatment of Renal Cell Carcinoma

Cancers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2471
Author(s):  
Ying Zhang ◽  
Jörg Ellinger ◽  
Manuel Ritter ◽  
Ingo G. H. Schmidt-Wolf
Keyword(s):  
Clinical Studies ◽  
Large Scale ◽  
Progression Free Survival ◽  
Preclinical Research ◽  
Cik Cells ◽  
Cell Immunotherapy ◽  
Integrated Therapy ◽  
Clinical Benefits ◽  
Cik Cell ◽  
Underlying Mechanisms

There is growing interest in cytokine-induced killer (CIK) cells on the integrated therapy of patients with RCC, especially those in the late stage or refractory to conventional chemotherapy and radiotherapy. In this review, a total of 15 clinical studies including 681 patients enrolled in CIK cell immunotherapy were outlined. Three-hundred-and-eighty-two patients with RCC were treated with CIK cells alone or in combination with DC vaccination, targeted agents sunitinib or sorafenib, and the PD-1 inhibitor pembrolizumab. Significantly improved 3-year overall survival rate was reported in four trials, whereas remarkably longer median progression-free survival was observed in three studies. Adverse reactions were mild and usually controllable fever and fatigue. Besides, preclinical research progresses were reviewed to increase our understanding about the underlying mechanisms of CIK cell cytotoxicity and identify potential targets to enhance their anti-tumor activity. These studies suggest that CIK cell-based immunotherapy has potential clinical benefits with a good safety profile and could become a promising approach in the combined therapies of RCC patients. However, further large-scale studies are required to evaluate the clinical efficacy of CIK cells and more efforts should be performed to identify the optimal CIK cell-based therapeutic regimen for RCC patients.

Optimization of a gmp-grade large-scale expansion protocol for cytokine-induced killer cells using gas-permeable static culture flasks

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S116
Author(s):  
P. Palmerini ◽  
E. Cappuzzello ◽  
A. Dalla Pietà ◽  
R. Sommaggio ◽  
G. Astori ◽  
...  
Keyword(s):  
Large Scale ◽  
Killer Cells ◽  
Static Culture ◽  
Expansion Protocol ◽  
Scale Expansion

scholarly journals Nanotopography reveals metabolites that maintain the immunosuppressive phenotype of mesenchymal stem cells

2019 ◽  
Author(s):  
Ewan A Ross ◽  
Lesley-Anne Turner ◽  
Anwar Saeed ◽  
Karl V Burgess ◽  
Gavin Blackburn ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Large Scale ◽  
Therapeutic Potential ◽  
Bioactive Metabolites ◽  
Multipotent Stem Cells ◽  
Immunomodulatory Capacity ◽  
Standard Culture ◽  
Scale Expansion ◽  
First Time

Mesenchymal stem cells (MSCs) are multipotent stem cells that are immunosuppressive and thus of considerable therapeutic potential in transplant operations. However, MSCs rapidly differentiate once in culture, making their large-scale expansion for use in immunosuppressive therapies challenging. Although the differentiation mechanisms of MSCs have been extensively investigated using materials, little is known about how materials can modulate paracrine activities of MSCs. Here, we show for the first time that nanotopography can control the immunomodulatory capacity of MSCs through decreased intracellular tension increasing oxidative glycolysis. We also use the nanotopography to identify bioactive metabolites that modulate intracellular tension, growth and immunomodulatory phenotype of MSCs in standard culture. Our findings show a novel route to support large-scale expansion of functional MSCs for therapeutic purposes

AGRICULTURAL PRODUCTION OF CA DONG PEOPLE IN RESETTLEMENT AREA OF TRANH RIVER HYDROPOWER NUMBER 2: CURRENT SITUATION AND IMPACT FACTORS

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Bùi Thị Bích Lan
Keyword(s):  
Social Relations ◽  
Agricultural Production ◽  
Large Scale ◽  
Transition Process ◽  
Impact Factors ◽  
Small Scale ◽  
Scale Production ◽  
Technical Application ◽  
Production Models ◽  
Role Of The State

In Vietnam, the construction of hydropower projects has contributed significantly in the cause of industrialization and modernization of the country. The place where hydropower projects are built is mostly inhabited by ethnic minorities - communities that rely primarily on land, a very important source of livelihood security. In the context of the lack of common productive land in resettlement areas, the orientation for agricultural production is to promote indigenous knowledge combined with increasing scientific and technical application; shifting from small-scale production practices to large-scale commodity production. However, the research results of this article show that many obstacles in the transition process are being posed such as limitations on natural resources, traditional production thinking or the suitability and effectiveness of scientific - technical application models. When agricultural production does not ensure food security, a number of implications for people’s lives are increasingly evident, such as poverty, preserving cultural identity, social relations and resource protection. Since then, it has set the role of the State in researching and building appropriate agricultural production models to exploit local strengths and ensure sustainability.

I. S. A. Baud. Forms of Production and Women's Labour: Gender Aspects of Industrialisation in India and Mexico. New Delhi: Sage Publications. 1992. 321 pp.Hardbound. Price: Indian Rs 295.00.

1993 ◽  
Vol 32 (1) ◽  
pp. 129-131
Author(s):  
Naureen Talha
Keyword(s):  
Large Scale ◽  
Indian Society ◽  
New Delhi ◽  
Scale Production ◽  
Female Labour ◽  
Self Employment ◽  
Commodity Production ◽  
Large Scale Production ◽  
Mexican Society ◽  
Small Production

The literature on female labour in Third World countries has become quite extensive. India, being comparatively more advanced industrially, and in view of its size and population, presents a pictures of multiplicity of problems which face the female labour market. However, the author has also included Mexico in this analytical study. It is interesting to see the characteristics of developing industrialisation in two different societies: the Indian society, which is conservative, and the Mexican society, which is progressive. In the first chapter of the book, the author explains that he is not concerned with the process of industrialisation and female labour employed at different levels of work, but that he is interested in forms of production and women's employment in large-scale production, petty commodity production, marginal small production, and self-employment in the informal sector. It is only by analysis of these forms that the picture of females having a lower status is understood in its social and political setting.

Solid Lipid Nanoparticles: A Promising Drug Delivery Technology

2009 ◽  
Vol 2 (2) ◽  
pp. 509-516
Author(s):  
S. Pragati ◽  
S. Kuldeep ◽  
S. Ashok ◽  
M. Satheesh
Keyword(s):  
Drug Delivery ◽  
Solid Lipid Nanoparticles ◽  
Large Scale ◽  
Colloidal Particles ◽  
Scale Up ◽  
Lipid Nanoparticles ◽  
Scale Production ◽  
Research Activity ◽  
New Approach ◽  
Solid Lipid

One of the situations in the treatment of disease is the delivery of efficacious medication of appropriate concentration to the site of action in a controlled and continual manner. Nanoparticle represents an important particulate carrier system, developed accordingly. Nanoparticles are solid colloidal particles ranging in size from 1 to 1000 nm and composed of macromolecular material. Nanoparticles could be polymeric or lipidic (SLNs). Industry estimates suggest that approximately 40% of lipophilic drug candidates fail due to solubility and formulation stability issues, prompting significant research activity in advanced lipophile delivery technologies. Solid lipid nanoparticle technology represents a promising new approach to lipophile drug delivery. Solid lipid nanoparticles (SLNs) are important advancement in this area. The bioacceptable and biodegradable nature of SLNs makes them less toxic as compared to polymeric nanoparticles. Supplemented with small size which prolongs the circulation time in blood, feasible scale up for large scale production and absence of burst effect makes them interesting candidates for study. In this present review this new approach is discussed in terms of their preparation, advantages, characterization and special features.

Stepwise Development of Biomimetic Chimeric Peptides for Gene Delivery

2020 ◽  
Vol 27 (8) ◽  
pp. 698-710
Author(s):  
Roya Cheraghi ◽  
Mahboobeh Nazari ◽  
Mohsen Alipour ◽  
Saman Hosseinkhani
Keyword(s):  
Gene Delivery ◽  
Targeted Delivery ◽  
Viral Vectors ◽  
Large Scale ◽  
Transfection Efficiency ◽  
Cationic Lipids ◽  
Target Cells ◽  
Scale Production ◽  
Stepwise Development ◽  
Chimeric Peptides

Gene-based therapy largely relies on the vector type that allows a selective and efficient transfection into the target cells with maximum efficacy and minimal toxicity. Although, genes delivered utilizing modified viruses transfect efficiently and precisely, these vectors can cause severe immunological responses and are potentially carcinogenic. A promising method of overcoming this limitation is the use of non-viral vectors, including cationic lipids, polymers, dendrimers, and peptides, which offer potential routes for compacting DNA for targeted delivery. Although non-viral vectors exhibit reduced transfection efficiency compared to their viral counterpart, their superior biocompatibility, non-immunogenicity and potential for large-scale production make them increasingly attractive for modern therapy. There has been a great deal of interest in the development of biomimetic chimeric peptides. Biomimetic chimeric peptides contain different motifs for gene translocation into the nucleus of the desired cells. They have motifs for gene targeting into the desired cell, condense DNA into nanosize particles, translocate the gene into the nucleus and enhance the release of the particle into the cytoplasm. These carriers were developed in recent years. This review highlights the stepwise development of the biomimetic chimeric peptides currently being used in gene delivery.

Sign in / Sign up

Export Citation Format

Share Document