Economics and quality attributes of hMSC production in xeno-free bioprocessing media

Human Mesenchymal Stem Cells (hMSCs) are key raw material in Regenerative Medicine and are widely used for therapeutics, engineered tissues, and medical devices. Yet, achieving an economical bioprocess for hMSC production remains a significant challenge for industry. Bioprocess economic modeling highlights media as a major cost driver in cell manufacturing. Hence, the availability of efficient and robust xeno-free bioprocessing media will not only reduce manufacturing cost, but also decrease regulatory burden associated with bovine serum components found in traditional culture media. Here, we evaluated and compared hMSCs quality parameters in bovine serum-containing and xeno-free bioprocess media formulations and assessed quality parameters such as cell identity, potency and functionality. Cells in xeno-free media maintained critical hMSC functional properties including angiogenic cytokine (FGF, HGF, IL8, TIMP1, TIMP2, and VEGF) secretion, trilineage differentiation, and immunomodulatory potential. In addition, hMSCs cultured in xeno-free media expanded rapidly and achieved confluency within 4-5 days of culture without media exchange. The economics of hMSC expansion in this xeno-free media were modeled and compared to other competitive hMSC cell or media systems where it consistently outperformed traditional hMSC systems by more than 8 fold on the critical productivity metric of Million cells per Liter, making it ideal for industrial-scale manufacturing of hMSCs

Peak MSC—Are We There Yet?

Human mesenchymal stem cells (hMSCs) are a critical raw material for many regenerative medicine products, including cell-based therapies, engineered tissues, or combination products, and are on the brink of radically changing how the world of medicine operates. Their unique characteristics, potential to treat many indications, and established safety profile in more than 800 clinical trials have contributed to their current consumption and will only fuel future demand. Given the large target patient populations with typical dose sizes of 10's to 100's of millions of cells per patient, and engineered tissues being constructed with 100's of millions to billions of cells, an unprecedented demand has been created for hMSCs. The fulfillment of this demand faces an uphill challenge in the limited availability of large quantities of pharmaceutical grade hMSCs for the industry—fueling the need for parallel rapid advancements in the biomanufacturing of this living critical raw material. Simply put, hMSCs are no different than technologies like transistors, as they are a highly technical and modular product that requires stringent control over manufacturing that can allow for high quality and consistent performance. As hMSC manufacturing processes are optimized, it predicts a future time of abundance for hMSCs, where scientists and researchers around the world will have access to a consistent and readily available supply of high quality, standardized, and economical pharmaceutical grade product to buy off the shelf for their applications and drive product development—this is “Peak MSC.

Advanced Hydrogen Transport Membranes for Vision 21 Fossil Fuel Plants

Seeks to economically eliminate the environmental concerns associated with the use of fossil fuels

Cost Effectiveness in River Management: Evaluation of Integrated River Policy System in Tidal Ouse

The River Ouse forms a significant part of Humber river system, which drains about one fifth the land area of England and provides the largest fresh water source to the North Sea from UK. The river quality in the tidal river suffered from sag of dissolved oxygen (DO) during last few decades, deteriorated by the effluent discharges. The Environment Agency (EA) proposed to increase the water quality of Ouse by implementing more potent environmental policies. This paper explores the cost effectiveness of water management in the Tidal Ouse through various options by taking into account the variation of assimilative capacity of river water, both in static and dynamic scope of time. Reduction in both effluent discharges and water abstraction were considered along side with choice of effluent discharge location. Different instruments of environmental policy, the emission tax-subsidy (ETS) scheme and tradable pollution permits (TPP) systems were compared with the direct quantitative control approach. This paper at the last illustrated an empirical example to reach a particular water quality target in the tidal Ouse at the least cost, through a solution of constrained optimisation problem. The results suggested significant improvement in the water quality with less cost than current that will fail the target in low flow year

Considerations and methods for placebo controls in surgical trials (ASPIRE guidelines)

Placebo comparisons are increasingly being considered for randomised trials assessing the efficacy of surgical interventions. The aim of this Review is to provide a summary of knowledge on placebo controls in surgical trials. A placebo control is a complex type of comparison group in the surgical setting and, although powerful, presents many challenges. This Review outlines what a placebo control entails and present understanding of this tool in the context of surgery. We consider when placebo controls in surgery are acceptable (and when they are desirable) in terms of ethical arguments and regulatory requirements, how a placebo control should be designed, how to identify and mitigate risk for participants in these trials, and how such trials should be done and interpreted. Use of placebo controls is justified in randomised controlled trials of surgical interventions provided there is a strong scientific and ethical rationale. Surgical placebos might be most appropriate when there is poor evidence for the efficacy of the procedure and a justified concern that results of a trial would be associated with high risk of bias, particularly because of the placebo effect. Feasibility work is recommended to optimise the design and implementation of randomised controlled trials. This Review forms an outline for best practice and provides guidance, in the form of the Applying Surgical Placebo in Randomised Evaluations (known as ASPIRE) checklist, for those considering the use of a placebo control in a surgical randomised controlled trial

Controlling myoblast phenotype with RGD -modified alginate matrices.

New strategies are being developed to grow tissues and organs for transplantation from cells and biomaterials. Biomaterials are critical components of engineered tissues, as they act as scaffolds for new tissues development. To date, however, engineered tissues do not have the complete structure or function of native tissues they are to replace. It is important to provide cells within these tissues with biological signals to help guide cell function during engineered tissue development. We hypothesized that the phenotype of cells adherent to a biomaterial could be regulated by controlling the mechanism of cell adhesion to the biomaterial, and we developed a model biomaterial system based on alginate hydrogels to address this hypothesis. Alginates are hydrophilic polysaccharides composed of mannuronic (M) and guluronic (G) acid monomers that gel in the presence of divalent cations such as Ca+2. We modified the alginates with RGD-peptides using carbodiimide chemistry. Ligand type and density may be varied on alginates of varying M:G over several orders of magnitude, with incorporation efficiency typically >60%. Myoblasts adhered, proliferated and differentiated on RGD-alginate hydrogels with a surface density of 10 fmols/cm2. Myoblast adhesion specificity was demonstrated, as soluble RGD (1mM) completely inhibited adhesion to the substrates, while RGE-peptides (1mM) had no effect. Myoblast function was controlled by varying ligand type, ligand density, and M:G of the alginate. Myoblast proliferation increased on RGD vs. YIGSR peptide ligands (density = 10 fmols/cm2), and proliferation and fusion increased by increasing RGD-density from 1--100 fmols/cm2. Furthermore, myoblast proliferation and fusion, but not muscle-specific gene expression, were dependent on the M:G ratio of alginates. Varying M:G ratio from 30:70 to 65:35 increased myoblast proliferation and fusion, but did not alter the expression of MyoD or myogenin (myoblast transcription factors) or the activity levels of creatine kinase. This effect was caused by calcium calcium ions released from different alginate substrates. In conclusion, myoblast phenotype may be controlled by varying ligand type and density at the material surface, and by modulating local calcium concentrations using different alginate types.Ph.D.Applied SciencesBiomedical engineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/126366/2/3016945.pd

Thiazolidinedione toxicity to isolated hepatocytes revealed by coherent multiprobe flourescence microscopy and correlated with multiparameter flow cytometry of peripheral leukocytes

Thiazolidinediones (TZDs) are effective for the treatment of adult-onset insulin-resistant diabetes. Unfortunately, TZDs are associated with sporadic hepatic dysfunction that is not predictable from experimental animal studies. We investigated the response of isolated rat and human hepatocytes to various TZDs using biochemical assays, coherent multiprobe fluorescence microscopy and flow cytometric analyses. The results identified direct effects of TZD on mitochondria from live human and rodent hepatocytes. The multiprobe fluorescence assays showed disruption of mitochondrial activity as an initiating event followed by increased membrane permeability, calcium influx and nuclear condensation. Other TZD-related cellular effects were increased hepatic enzyme leakage, decreased reductive metabolism and cytoplasmic adenosine triphosphate depletion. Mitochondrial effects were similar in cryopreserved hepatocytes from diabetic or non-diabetic donors. Peripheral blood mononuclear cells (PBMCs) had baseline mitochondrial energetics and metabolism comparable with isolated hepatocytes. Mitochondrial effects in isolated hepatocytes were found in human PBMCs exposed to the TZDs. The relative potency of TZDs for causing hepatocyte and PBMC effects was troglitazone >pioglitazone >rosiglitazone. These studies clearly demonstrated that hepatic alterations in vitro are characteristic of TZDs, with only quantitative differences in subcellular organelle dysfunction. Monitoring mitochondrial function in isolated PBMCs may be beneficial in diabetics undergoing TZD therapy.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41920/1/204-75-7-425_s002040100251.pd