scholarly journals Automated Large-Scale Production of Paclitaxel Loaded Mesenchymal Stromal Cells for Cell Therapy Applications

Pharmaceutics ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 411 ◽  
Author(s):  
Daniela Lisini ◽  
Sara Nava ◽  
Simona Frigerio ◽  
Simona Pogliani ◽  
Guido Maronati ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Large Scale ◽  
Good Manufacturing Practice ◽  
Scale Production ◽  
Large Scale Production ◽  
Advanced Therapies ◽  
Oncological Diseases

Mesenchymal stromal cells (MSCs) prepared as advanced therapies medicinal products (ATMPs) have been widely used for the treatment of different diseases. The latest developments concern the possibility to use MSCs as carrier of molecules, including chemotherapeutic drugs. Taking advantage of their intrinsic homing feature, MSCs may improve drugs localization in the disease area. However, for cell therapy applications, a significant number of MSCs loaded with the drug is required. We here investigate the possibility to produce a large amount of Good Manufacturing Practice (GMP)-compliant MSCs loaded with the chemotherapeutic drug Paclitaxel (MSCs-PTX), using a closed bioreactor system. Cells were obtained starting from 13 adipose tissue lipoaspirates. All samples were characterized in terms of number/viability, morphology, growth kinetics, and immunophenotype. The ability of MSCs to internalize PTX as well as the antiproliferative activity of the MSCs-PTX in vitro was also assessed. The results demonstrate that our approach allows a large scale expansion of cells within a week; the MSCs-PTX, despite a different morphology from MSCs, displayed the typical features of MSCs in terms of viability, adhesion capacity, and phenotype. In addition, MSCs showed the ability to internalize PTX and finally to kill cancer cells, inhibiting the proliferation of tumor lines in vitro. In summary our results demonstrate for the first time that it is possible to obtain, in a short time, large amounts of MSCs loaded with PTX to be used in clinical trials for the treatment of patients with oncological diseases.

scholarly journals Large-scale production of megakaryocytes from human pluripotent stem cells by chemically defined forward programming

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Thomas Moreau ◽  
Amanda L. Evans ◽  
Louella Vasquez ◽  
Marloes R. Tijssen ◽  
Ying Yan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Blood Platelets ◽  
Basic Research ◽  
Good Manufacturing Practice ◽  
Human Pluripotent Stem Cells ◽  
Scale Production ◽  
Large Scale Production

Abstract The production of megakaryocytes (MKs)—the precursors of blood platelets—from human pluripotent stem cells (hPSCs) offers exciting clinical opportunities for transfusion medicine. Here we describe an original approach for the large-scale generation of MKs in chemically defined conditions using a forward programming strategy relying on the concurrent exogenous expression of three transcription factors: GATA1, FLI1 and TAL1. The forward programmed MKs proliferate and differentiate in culture for several months with MK purity over 90% reaching up to 2 × 105 mature MKs per input hPSC. Functional platelets are generated throughout the culture allowing the prospective collection of several transfusion units from as few as 1 million starting hPSCs. The high cell purity and yield achieved by MK forward programming, combined with efficient cryopreservation and good manufacturing practice (GMP)-compatible culture, make this approach eminently suitable to both in vitro production of platelets for transfusion and basic research in MK and platelet biology.

scholarly journals Inhibition of Human Malignant Pleural Mesothelioma Growth by Mesenchymal Stromal Cells

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1427
Author(s):  
Valentina Coccè ◽  
Silvia La Monica ◽  
Mara Bonelli ◽  
Giulio Alessandri ◽  
Roberta Alfieri ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Mesenchymal Stromal Cells ◽  
Malignant Pleural Mesothelioma ◽  
Stromal Cells ◽  
Large Scale ◽  
Significant Inhibition ◽  
G1 Arrest ◽  
Pleural Mesothelioma ◽  
Scale Production

Background: Malignant Pleural Mesothelioma (MPM) is an aggressive tumor that has a significant incidence related to asbestos exposure with no effective therapy and poor prognosis. The role of mesenchymal stromal cells (MSCs) in cancer is controversial due to their opposite effects on tumor growth and in particular, only a few data are reported on MSCs and MPM. Methods: We investigated the in vitro efficacy of adipose tissue-derived MSCs, their lysates and secretome against different MPM cell lines. After large-scale production of MSCs in a bioreactor, their efficacy was also evaluated on a human MPM xenograft in mice. Results: MSCs, their lysate and secretome inhibited MPM cell proliferation in vitro with S or G0/G1 arrest of the cell cycle, respectively. MSC lysate induced cell death by apoptosis. The efficacy of MSC was confirmed in vivo by a significant inhibition of tumor growth, similar to that produced by systemic administration of paclitaxel. Interestingly, no tumor progression was observed after the last MSC treatment, while tumors started to grow again after stopping chemotherapeutic treatment. Conclusions: These data demonstrated for the first time that MSCs, both through paracrine and cell-to-cell interaction mechanisms, induced a significant inhibition of human mesothelioma growth. Since the prognosis for MPM patients is poor and the options of care are limited to chemotherapy, MSCs could provide a potential new therapeutic approach for this malignancy.

scholarly journals Inactivated Platelet Lysate Supports Proliferation and Immunomodulant Characteristics Of Mesenchymal Stromal Cells in GMP Culture Condition

2020 ◽  
Author(s):  
Katia Mareschi ◽  
Sara Castiglia ◽  
Aloe Adamini ◽  
Deborah Rustichelli ◽  
Elena Marini ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Good Alternative ◽  
Good Manufacturing Practice ◽  
Platelet Lysate ◽  
T Cell Subsets ◽  
Cytokine Release ◽  
Pathogen Inactivation ◽  
Advanced Therapy

For their clinical use Mesenchymal Stromal Cells (MSCs), isolated from bone marrow (BM-MSCs) are considered Advanced Therapy Medicinal Products (ATMP) and need to be produced according to Good Manufacturing Practice (GMP). Human platelet lysate (HPL) represents a good GMP-compliant alternative to animal serum and after pathogen inactivation with Psoralen was more efficient and safer to produce MSCs in GMP. In this study MSCs cultivated in FBS (FBS-MSC) or inactivated HPL (iHPL-MSC), were compared for their immunomodulant properties. In particular, the effects of MSCs on: 1)proliferation of total Lymphocytes (Ly) and on naïve T Ly subsets induced to differentiate versus Th1 and Th2 Ly; 2) the immunophenotype of different T cell subsets; 3)the cytokine release to verify Th1, Th2 and Th17 polarization were analyzed by using in vitro co-culture system. We observed that iHPL-MSCs showed the same immunomodulant properties observed in the FBS-MSCs co-cultures. Although, a more efficient effect on the increase of naïve T cells and, in the Th1 cytokine release related to iHPL was observed. This study confirms that iHPL, used as medium supplement, may be considered a good alternative to FBS for a GMP-compliant MSC expansion to preserve their immunomodulant proprieties.

Optimizing in vitro large scale production of giant reed (Arundo donax L.) by liquid medium culture

2014 ◽  
Vol 69 ◽  
pp. 21-27 ◽  
Author(s):  
Valeria Cavallaro ◽  
Cristina Patanè ◽  
Salvatore L. Cosentino ◽  
Isabella Di Silvestro ◽  
Venera Copani
Keyword(s):  
Liquid Medium ◽  
Large Scale ◽  
Arundo Donax ◽  
Giant Reed ◽  
Scale Production ◽  
Large Scale Production ◽  
Medium Culture ◽  
Arundo Donax L

Large-scale production of polyoma middle T antigen by using genetically engineered tumors

1985 ◽  
Vol 5 (7) ◽  
pp. 1795-1799
Author(s):  
D R Kaplan ◽  
B Bockus ◽  
T M Roberts ◽  
J Bolen ◽  
M Israel ◽  
...  
Keyword(s):  
Nude Mice ◽  
Large Scale ◽  
T Antigen ◽  
Genetically Engineered ◽  
Scale Production ◽  
Large Scale Production ◽  
Metallothionein Promoter ◽  
Polyoma Middle T Antigen ◽  
Nih 3T3

A recombinant plasmid containing a metallothionein promoter-polyoma middle T cDNA fusion was constructed and used to transfect NIH 3T3 cells. Transformed cells expressing middle T were injected into nude mice. Within 3 weeks, each mouse produced tumors containing middle T equivalent to that in 250 to 1,000 100-mm dishes of polyomavirus-infected cells. This middle T, partially purified by immunoaffinity chromatography, retained activity as measured by its ability to be phosphorylated in vitro. The combined approach of fusing strong promoters to genes of interest and utilizing nude mice to grow large quantities of cells expressing the gene provides a quick, inexpensive alternative to other expression systems.

scholarly journals Production of Phytotoxic Metabolite Using Biphasic Fermentation System from Strain C1136 of Lasiodiplodia pseudotheobromae, a Potential Bioherbicidal Agent

2017 ◽  
Vol 9 (3) ◽  
pp. 371-377
Author(s):  
Charles Oluwaseun ADETUNJI ◽  
Julius Kola OLOKE ◽  
Gandham PRASAD ◽  
Moses ABALAKA ◽  
Emenike Onyebum IROKANULO
Keyword(s):  
Large Scale ◽  
Wild Strain ◽  
Fermentation Medium ◽  
Scale Production ◽  
Conventional Farming ◽  
Large Scale Production ◽  
Phytotoxic Metabolite ◽  
Biphasic Fermentation

Formulation of effective and environmental friendly bioherbicides depends on the type of fermentation medium used for the production of phytotoxic metabolites. The effect of biomass, colony forming unit and the phytotoxic metabolite produced from the biphasic fermentation was carried out, while the phytotoxic metabolite was  tested in vivo and in-vitro on Echinochola crus-galli and dicotyledonous Chromolaena odorata. The mutant strain of Lasiodiplodia pseudotheobromae C1136 (Lp90) produced the highest amount of conidia and the largest necrotic area on the two tested weeds when compared to its wild strain in the different biphasic media combinations. The study revealed that the biphasic system containing PDB + rice produced the highest bioherbicidal activities. Therefore, the phytotoxic metabolites from strain C1136 are suggested for large scale production of bioherbicides for the management of weeds in conventional farming to improve yield and enhance food security.

scholarly journals Reproducible Large-Scale Isolation of Exosomes from Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells and Their Application in Acute Kidney Injury

2020 ◽  
Vol 21 (13) ◽  
pp. 4774 ◽  
Author(s):  
Jun Ho Lee ◽  
Dae Hyun Ha ◽  
Hyeon-kyu Go ◽  
Jinkwon Youn ◽  
Hyun-keun Kim ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Adipose Tissue ◽  
Stromal Cells ◽  
Large Scale ◽  
Kidney Diseases ◽  
Kidney Injury ◽  
Preclinical Studies ◽  
Scale Production ◽  
Positive Effects ◽  
Large Scale Production

Acute kidney injury (AKI) is a fatal medical episode caused by sudden kidney damage or failure, leading to the death of patients within a few hours or days. Previous studies demonstrated that exosomes derived from various mesenchymal stem/stromal cells (MSC-exosomes) have positive effects on renal injuries in multiple experimental animal models of kidney diseases including AKI. However, the mass production of exosomes is a challenge not only in preclinical studies with large animals but also for successful clinical applications. In this respect, tangential flow filtration (TFF) is suitable for good manufacturing practice (GMP)-compliant large-scale production of high-quality exosomes. Until now, no studies have been reported on the use of TFF, but rather ultracentrifugation has been almost exclusively used, to isolate exosomes for AKI therapeutic application in preclinical studies. Here, we demonstrated the reproducible large-scale production of exosomes derived from adipose tissue-derived MSC (ASC-exosomes) using TFF and the lifesaving effect of the ASC-exosomes in a lethal model of cisplatin-induced rat AKI. Our results suggest the possibility of large-scale stable production of ASC-exosomes without loss of function and their successful application in life-threatening diseases.

scholarly journals Large-Scale Production of Lentiviral Vectors: Current Perspectives and Challenges

Pharmaceutics ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1051
Author(s):  
Eduardo Martínez-Molina ◽  
Carlos Chocarro-Wrona ◽  
Daniel Martínez-Moreno ◽  
Juan A. Marchal ◽  
Houria Boulaiz
Keyword(s):  
Viral Vectors ◽  
Large Scale ◽  
Lentiviral Vectors ◽  
Good Manufacturing Practice ◽  
Scale Production ◽  
Gene Therapies ◽  
Packaging Cell ◽  
Large Scale Production ◽  
Target Production

Lentiviral vectors (LVs) have gained value over recent years as gene carriers in gene therapy. These viral vectors are safer than what was previously being used for gene transfer and are capable of infecting both dividing and nondividing cells with a long-term expression. This characteristic makes LVs ideal for clinical research, as has been demonstrated with the approval of lentivirus-based gene therapies from the Food and Drug Administration and the European Agency for Medicine. A large number of functional lentiviral particles are required for clinical trials, and large-scale production has been challenging. Therefore, efforts are focused on solving the drawbacks associated with the production and purification of LVsunder current good manufacturing practice. In recent years, we have witnessed the development and optimization of new protocols, packaging cell lines, and culture devices that are very close to reaching the target production level. Here, we review the most recent, efficient, and promising methods for the clinical-scale production ofLVs.

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