Identification of Five Developmental Processes during Chondrogenic Differentiation of Embryonic Stem Cells

Chondrogenesis is the complex process that leads to the establishment of cartilage and bone formation. Due to their ability to differentiate in vitro and mimic development, embryonic stem cells (ESCs) show great potential for investigating developmental processes. In this study, we used chondrogenic differentiation of ESCs as a model to analyze morphogenetic events during chondrogenesis.ESCs were differentiated into the chondrocyte lineage, forming small cartilaginous aggregates in suspension. Differentiated ESCs showed that chondrogenesis was typically characterized by five overlapping stages. During the first stage, cell condensation and aggregate formation was observed. The second stage was characterized by differentiation into chondrocytes and fibril scaffold formation within spherical aggregates. Deposition of cartilaginous extracellular matrix and cartilage formation were hallmarks of the third stage. Apoptosis of chondrocytes, hypertrophy and/or degradation of cartilage occurred during the fourth stage. Finally, during the fifth stage, bone replacement with membranous calcified tissues took place.We demonstrate that ESCs show the chondrogenic differentiation pathway from the pluripotent stem cell to terminal skeletogenesis through these five stages in vitro. During each stage, morphological changes acquired in preceding stages played an important role in further development as a scaffold or template in subsequent stages. The study of chondrogenesis via ESC differentiation may be informative to our further understanding of skeletal growth and regeneration

Derivation of Chondrogenically-Committed Cells from Human Embryonic Cells for Cartilage Tissue Regeneration

Background: Heterogeneous and uncontrolled differentiation of human embryonic stem cells (hESCs) in embryoid bodies (EBs) limits the potential use of hESCs for cell-based therapies. More efficient strategies are needed for the commitment and differentiation of hESCs to produce a homogeneous population of specific cell types for tissue regeneration applications. Methodology/Principal Findings: We report here that significant chondrocytic commitment of feeder-free cultured human embryonic stem cells (FF-hESCs), as determined by gene expression and immunostaining analysis, was induced by coculture with primary chondrocytes. Furthermore, a dynamic expression profile of chondrocyte-specific genes was observed during monolayer expansion of the chondrogenically-committed cells. Chondrogenically-committed cells synergistically responded to transforming growth factor-b1 (TGF-b1) and b1-integrin activating antibody by increasing tissue mass in pellet culture. In addition, when encapsulated in hydrogels, these cells formed cartilage tissue both in vitro and in vivo. In contrast, the absence of chondrocyte co-culture did not result in an expandable cell population from FF-hESCs. Conclusions/Significance: The direct chondrocytic commitment of FF-hESCs can be induced by morphogenetic factor

Detection of anthropogenic climate change using a fingerprint method

A fingerprint method for detecting anthropogenic climate change is applied to new simulations with a coupled ocean-atmosphere general circulation model (CGCM) forced by increasing concentrations of greenhouse gases and aerosols covering the years 1880 to 2050. In addition to the anthropogenic climate change signal, the space-time structure of the natural climate variability for near-surface temperatures is estimated from instrumental data over the last 134 years and two 1000 year simulations with CGCMs. The estimates are compared with paleoclimate data over 570 years. The space-time information on both the signal and the noise is used to maximize the signal-to-noise ratio of a detection variable obtained by applying an optimal filter (fingerprint) to the observed data. The inclusion of aerosols slows the predicted future warming. The probability that the observed increase in near-surface temperatures in recent decades is of natural origin is estimated to be less than 5%. However, this number is dependent on the estimated natural variability level, which is still subject to some uncertainty

Managing cooperative research and development: Partner selection and contract design

This paper explores the influence of partner selection and the design of contracts and agreements on the chances for a collaborative R&D venture to 'succeed'. It is based on questionnaire data regarding 49 such ventures undertaken by companies based in four Nordic countries. The age distribution of the sample is taken into account through use of statistical event history analysis, testing hypotheses regarding the determinants of the hazard rate. The results show that prior contacts with prospective partners improve the chances that a cooperation will succeed. Such contacts help prevent difficulties due to differences in corporate culture, inadequate technical capabilities, incongruous strategic intentions, etc. The analysis also suggests that contracts and agreements should avoid detailed specifications of the procedures to be followed during implementation. Such stipulations create problems by reducing the flexibility required in inherently uncertain R&D collaborations. Granting relative resource self-sufficiency and managerial autonomy to the venture organisation improves its chances of success