ABSTRACT
Aims: To investigate the Mongolian plant M. mohileviensis Down. for the presence of possible bioactive products that could be related to the traditional use of the plant in Mongolia. Methodology: Ethanolic and water extracts of both seeds and herb were tested for antiinflammatory and DPPH scavenging activity. Polysaccharides were isolated from the seeds using sequence of chromatographic methods. The polysaccharide fraction from the cold water extract was then analyzed for the presence of monosaccharides and their type of linkages by GC and GC-MS. The effect of the polysaccharides on the complement system was then determined. Results: The ethanolic and aqueous extracts of seeds and herbs, as well as crude polysaccharides from cold and hot water extracts exhibited a significant anti-inflammatory activity in the model based on histamine-induced paw edema. Anti-inflammatory effects of all samples were high during the 3rd hour of inflammation. Moreover, the ethanolic extracts of seeds and herbs exhibited DPPH scavenging activity. Phytochemical studies of the cold water seed polysaccharide revealed the presence of an uncommon pectic type polysaccharide. Galacturonic acid (38%) and rhamnose (30%) were present as the main monosaccharides, and linkage analyses revealed that galacturonic acid was present as terminal, 14 and 13,4 linked units and rhamnose basically as 13 linked units. The complement fixation activity was appr.15 μg/ml, substantially lower than that of the standard used. The results also indicate strongly that the M. mohileviensis and M. verticillata are two different Malva species.
ABSTRACT
Future clinical applications of human embryonic stem (hES) cells will require high-yield culture protocols. Currently, hES cells are mainly cultured in static tissue plates, which offer a limited surface and require repeated sub-culturing. Here we describe a stirred system with commercial dextran-based microcarriers coated with denatured collagen to scale-up hES cell production. Maintenance of pluripotency in the microcarrier-based stirred system was shown by immunocytochemical and flow cytometry analyses for pluripotency-associated markers. The formation of cavitated embryoid bodies expressing markers of endoderm, ectoderm and mesoderm was further evidence of maintenance of differentiation capability. Cell yield per volume of medium spent was more than 2-fold higher than in static plates, resulting in a significant decrease in cultivation costs. A total of 10(8) karyotypically stable hES cells were obtained from a unitary small vessel that needed virtually no manipulation during cell proliferation, decreasing risks of contamination. Spinner flasks are available up to working volumes in the range of several liters. If desired, samples from the homogenous suspension can be withdrawn to allow process validation needed in the last expansion steps prior to transplantation. Especially when thinking about clinical trials involving from dozens to hundreds of patients, the use of a small number of larger spinners instead of hundreds of plates or flasks will be beneficial. To our knowledge, this is the first description of successful scale-up of feeder- and Matrigel-free production of undifferentiated hES cells under continuous agitation, which makes this system a promising alternative for both therapy and research needs.