Serum and supplement optimization for EU GMP‐compliance in cardiospheres cell culture

Cardiac progenitor cells (CPCs) isolated as cardiospheres (CSs) and CS-derived cells (CDCs) are a promising tool for cardiac cell therapy in heart failure patients, having CDCs already been used in a phase I/II clinical trial. Culture standardization according to Good Manufacturing Practices (GMPs) is a mandatory step for clinical translation. One of the main issues raised is the use of xenogenic additives (e.g. FBS, foetal bovine serum) in cell culture media, which carries the risk of contamination with infectious viral/prion agents, and the possible induction of immunizing effects in the final recipient. In this study, B27 supplement and sera requirements to comply with European GMPs were investigated in CSs and CDCs cultures, in terms of process yield/efficiency and final cell product gene expression levels, as well as phenotype. B27− free CS cultures produced a significantly reduced yield and a 10-fold drop in c-kit expression levels versus B27+ media. Moreover, autologous human serum (aHS) and two different commercially available GMP AB HSs were compared with standard research-grade FBS. CPCs from all HSs explants had reduced growth rate, assumed a senescent-like morphology with time in culture, and/or displayed a significant shift towards the endothelial phenotype. Among three different GMP gamma-irradiated FBSs (giFBSs) tested, two provided unsatisfactory cell yields, while one performed optimally, in terms of CPCs yield/phenotype. In conclusion, the use of HSs for the isolation and expansion of CSs/CDCs has to be excluded because of altered proliferation and/or commitment, while media supplemented with B27 and the selected giFBS allows successful EU GMP-complying CPCs culture.     

Microbiological condition and shelf life of meat from hunted game birds

Hunted game birds (eight partridges, nine wood pigeons, 25 quails, 16 chilled and 16 frozen–thawed pheasants) were processed according to “Good Manufacturing Practice” rules. Microflora of skin, intestinal content and meat cuts (breast and thigh, both fresh and stored in vacuum package) was analysed. Listeria monocytogenes, Salmonella sp. and Campylobacter sp. were not recovered from any sample. Log microbial numbers on skin or in intestines were not significantly related to those on meat cuts. With the exception of pigeons, microbial numbers of the two meat cuts did not differ significantly (p > 0.05), and no significant increase in microbial numbers in vacuum-stored meat was found; the same applied to frozen–thawed compared to chilled pheasants. On meat, average total viable counts were <4.00 log cfu/cm² with a maximum of 6.48 log cfu/cm². Median Escherichia coli numbers were <2.00 log cfu/cm², maximum was 4.48 log cfu/cm². Meat cuts obtained from partridges, quails and pheasants demonstrated a shelf life of 1 week, provided they were kept vacuum-packaged at 0°C to 1°C.     

Stem cell cultivation in bioreactors

Cell-based therapies have generated great interest in the scientific and medical communities, and stem cells in particular are very appealing for regenerative medicine, drug screening and other biomedical applications. These unspecialized cells have unlimited self-renewal capacity and the remarkable ability to produce mature cells with specialized functions, such as blood cells, nerve cells or cardiac muscle. However, the actual number of cells that can be obtained from available donors is very low. One possible solution for the generation of relevant numbers of cells for several applications is to scale-up the culture of these cells in vitro. This review describes recent developments in the cultivation of stem cells in bioreactors, particularly considerations regarding critical culture parameters, possible bioreactor configurations, and integration of novel technologies in the bioprocess development stage. We expect that this review will provide updated and detailed information focusing on the systematic production of stem cell products in compliance with regulatory guidelines, while using robust and cost-effective approaches.     

Standardizing CAR-T therapy: Getting it scaled up

CAR-T therapy, grafting the specificity of a monoclonal antibody onto a T cell to target certain cancer cells, has been recognized as a promising therapeutic approach for cancer control as evidenced by the two CAR-T products proved by FDA in 2017. However, the unique heterogeneity of CAR-T therapy has restricted its production in a limited number of institutions and made it a boutique oncotherapy. By reviewing outstanding issues surrounding the commercial scale production of CAR-T therapy, we conclude that achieving mass production of CAR-T therapy without sacrificing its personalized nature is a worldwild challenge for making CAR-T a key element in the next generation of precision medicine, which can be achieved by standardizing 7 prominent factors that collectively determine the scale of CAR-T manufacturing.     

Viability of Bioprinted Cellular Constructs Using a Three Dispenser Cartesian Printer

Tissue engineering has centralized its focus on the construction of replacements for non-functional or damaged tissue. The utilization of three-dimensional bioprinting in tissue engineering has generated new methods for the printing of cells and matrix to fabricate biomimetic tissue constructs. The solid freeform fabrication (SFF) method developed for three-dimensional bioprinting uses an additive manufacturing approach by depositing droplets of cells and hydrogels in a layer-by-layer fashion. Bioprinting fabrication is dependent on the specific placement of biological materials into three-dimensional architectures, and the printed constructs should closely mimic the complex organization of cells and extracellular matrices in native tissue. This paper highlights the use of the Palmetto Printer, a Cartesian bioprinter, as well as the process of producing spatially organized, viable constructs while simultaneously allowing control of environmental factors. This methodology utilizes computer-aided design and computer-aided manufacturing to produce these specific and complex geometries. Finally, this approach allows for the reproducible production of fabricated constructs optimized by controllable printing parameters.     

Differential Sensitivity of Human Hepatocellular Carcinoma Xenografts to an IGF-II Neutralizing Antibody May Involve Activated STAT3

Hepatocellular carcinoma (HCC) is highly refractory to current therapeutics used in the clinic. DX-2647, a recombinant human antibody, potently neutralizes the action of insulin-like growth factor-II (IGF-II), a ligand for three cell-surface receptors (IGF-IR, insulin receptor A and B isoforms, and the cation-independent mannose-6-phosphate receptor) which is overexpressed in primary human HCC. DX-2647 impaired the growth of tumor xenografts of the HCC cell line, Hep3B; however, xenografts of the HCC cell line, HepG2, were largely unresponsive to DX-2647 treatment. Analysis of a number of aspects of the IGF signaling axis in both cell lines did not reveal any significant differences between the two. However, while DX-2647 abolished phospho (p)-IGF-IR, p-IR and p-AKT signaling in both cell lines, HepG2 showed high levels of p-STAT3, which was unaffected by DX-2647 treatment and was absent from the Hep3B cell line. The driver of p-STAT3 was found to be a secreted cytokine, and treatment of HepG2 cells with a pan- JAK kinase inhibitor resulted in a loss of p-STAT3. These findings implicate the activation of STAT3 as one pathway that may mediate resistance to IGF-II–targeted therapy in HCC.     

栀子黄色素的提制及应用研究

栀子黄色素是从栀子(Gardenia Jasminoides Ellis)植物的果实中,采用水浸提、醇沉淀的萃取工艺,经真空蒸发浓缩和喷雾干燥生产的安全性商、着色力强、色泽鲜艳、稳定性好、而且应用范围广、成本较低的天然黄色素,是食品工业上良好的着色剂。     

Social sustainability in the supply chain: Construct development and measurement validation

Research on social sustainability in developing countries has recently gained importance for both academics and practitioners. Studies in the supply chain management field take either a supplier or a manufacturer perspective that address predominantly corporate social responsibility (CSR) issues referring to the internal stakeholders. Our research integrates the literature on supplier, manufacturer, and customer responsibility and proposes the concept of supply chain social sustainability (SCSS) that refers to addressing social issues within the overall (upstream and downstream) supply chain. Furthermore, we develop and empirically validate scales for measuring SCSS using in-depth interviews and a survey in the Indian manufacturing industry. Our results suggest that SCSS consists of six underlying dimensions, namely equity, safety, health and welfare, philanthropy, ethics, human rights, in a 20-item valid and reliable scale. We discuss the implications of the findings for research and practice and suggest future research avenues.     

Dating furniture and coopered vessels without waney edge – Reconstructing historical wood-working in Austria with the help of dendrochronology

In the present study, 208 furniture and 168 coopered vessels from three Austrian museums were examined. Dendrochronology was used to date objects and to extract further information such as the necessary time for seasoning, wood loss through wood-working and methods of construction. In most cases sampling was done by sanding the cross section and making digital photographs using a picture frame and measuring digitally.The dendrochronological dates of the sampled furniture range between 1524 and 1937. The group of furniture includes cupboards, chests, tables, benches, commodes and beds. In many cases furniture was artfully painted and sometimes even shows a painted year. With the help of dendrochronology it was proved that some objects had been painted for some time after construction, or had been over-painted. Most furniture, however, was painted immediately after completion. In this case, the seasoning and storage time of the boards and the wood loss due to shaping can be verified. As an average value, 14 years have passed between the dendrochronological date of the outermost ring and the painting. The time span includes time of seasoning and storage and the rings lost by wood-working. This leads, on the one hand to a short storage time of less than 10 years and on the other hand to very little wood loss due to manufacturing. Those boards being less shaped turned out to be back panels of cupboards, therefore they are recommended to be sampled for dating.Coopered vessels were dated between 1612 and 1940. There was evidence that staves were split and not sawn in many cases. The staves were often split out of the outermost part of the tree and hardly any wood was worked away which was proved by the close dendrochronological dates of the single staves of a vessel.Since there is a short time of storage and only little wood loss through wood-working, dating of objects without a waney edge becomes reasonable.     

朝鲜半岛旧石器时代晚期的有柄尖刃器

有柄尖刃器是朝鲜半岛旧石器时代晚期代表性狩猎工具,1964年在公州石壮里遗址中首次发现。原料多选择硅质页岩或与之相似的石料,多见于25,000-15,000 BP之间。之后随着石镞等复合工具的出现,有柄尖刃器逐渐被取代。本文观察和研究的305件有柄尖刃器来自朝鲜半岛33个遗址,通过分析其形态属性及工业类型,揭示有柄尖刃器的制作技法,探讨其与相邻地区的关系。在中国旧石器时代遗存中虽然也有关于有柄尖刃器的零星报道,但鲜有确凿者,望本文能为中国的相关研究提供有价值的参考资料。