Design of dual peptide-conjugated hydrogels for proliferation and differentiation of human pluripotent stem cells.

Completely synthetic cell cultivation materials for human pluripotent stem cells (hPSCs) are important for the future clinical use of hPSC-derived cells. Currently, cell culture materials conjugated with extracellular matrix (ECM)-derived peptides are being prepared using only one specific integrin-targeting peptide. We designed dual peptide-conjugated hydrogels, for which each peptide was selected from different ECM sites: the laminin ß4 chain and fibronectin or vitronectin, which can target α6ß1 and α2ß1 or αVß5. hPSCs cultured on dual peptide-conjugated hydrogels, especially on hydrogels conjugated with peptides obtained from the laminin ß4 chain and vitronectin with a low peptide concentration of 200 µg/mL, showed high proliferation ability over the long term and differentiated into cells originating from 3 germ layers in vivo as well as a specific lineage of cardiac cells. The design of grafting peptides was also important, for which a joint segment and positive amino acids were added into the designed peptide. Because of the designed peptides on the hydrogels, only 200 µg/mL peptide solution was sufficient for grafting on the hydrogels, and the hydrogels supported hPSC cultures long-term; in contrast, in previous studies, greater than 1000 µg/mL peptide solution was needed for the grafting of peptides on cell culture materials.

Xeno-free culture and proliferation of hPSCs on 2D biomaterials.

Human pluripotent stem cells (human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs)) have unlimited proliferative potential, whereas adult stem cells such as bone marrow-derived stem cells and adipose-derived stem cells have problems with aging. When hPSCs are intended to be cultured on feeder-free or xeno-free conditions without utilizing mouse embryonic fibroblasts or human fibroblasts, they cannot be cultured on conventional tissue culture polystyrene dishes, as adult stem cells can be cultured but should be cultivated on material surfaces grafted or coated with (a) natural or recombinant extracellular matrix (ECM) proteins, (b) ECM protein-derived peptides and specific synthetic polymer surfaces in xeno-free and/or chemically defined conditions. This review describes current developing cell culture biomaterials for the proliferation of hPSCs while maintaining the pluripotency and differentiation potential of the cells into 3 germ layers. Biomaterials for the cultivation of hPSCs without utilizing a feeder layer are essential to decrease the risk of xenogenic molecules, which contributes to the potential clinical usage of hPSCs. ECM proteins such as human recombinant vitronectin, laminin-511 and laminin-521 have been utilized instead of Matrigel for the feeder-free cultivation of hPSCs. The following biomaterials are also discussed for hPSC cultivation: (a) decellularized ECM, (b) peptide-grafted biomaterials derived from ECM proteins, (c) recombinant E-cadherin-coated surface, (d) polysaccharide-immobilized surface, (e) synthetic polymer surfaces with and without bioactive sites, (f) thermoresponsive polymer surfaces with and without bioactive sites, and (g) synthetic microfibrous scaffolds.

Emissions of polycyclic aromatic hydrocarbons from batch hot mix asphalt plants.

This study was set out to assess the characteristics of polycyclic aromatic hydrocarbon (PAH) emissions from batch hot mix asphalt (HMA) plants and PAH removal efficiencies associated with their installed air pollution control devices. Field samplings were conducted on six randomly selected batch HMA plants. For each selected plant, stack flue gas samples were collected from both stacks of the batch mixer (n = 5) and the preheating boiler (n = 5), respectively. PAH samples were also collected from the field to assess PAHs that were directly emitted from the discharging chute (n = 3). To assess PAH removal efficiencies of the installed air pollution control devices, PAH contents in both cyclone fly ash (n=3) and bag filter fly ash (n = 3) were analyzed. Results show that the total PAH concentration (mean; RSD) in the stack flue gas of the batch mixer (354 microg/Nm3; 78.5%) was higher than that emitted from the discharging chute (107 microg/Nm3; 70.1%) and that in the stack flue gas of the preheating boiler (83.7 microg/Nm3; 77.6%). But the total BaPeq concentration of that emitted from the discharging chute (0.950 microg/Nm3; 84.4%) was higher than contained in the stack flue gas of the batch mixer (0.629 microg/Nm3; 86.8%) and the stack flue gas of the preheating boiler (= 0.112 microg/Nm3; 80.3%). The mean total PAH emission factor for all selected batch mix plants (= 139 mg/ton x product) was much higher than that reported by U.S. EPA for the drum mix asphalt plant (range = 11.8-79.0 mg/ton x product). We found the overall removal efficiency of the installed air pollution control devices (i.e., cyclone + bag filter) on total PAHs and total BaPeq were 22.1% and 93.7%, respectively. This implies that the installed air pollution control devices, although they have a very limited effect on the removal of total PAHs, do significantly reduce the carcinogenic potencies associated with PAH emissions from batch HMA plants.