Background and Objective. It is important to guarantee the quality of stem cells. Serial passage is the main approach to expand stem cells. This study evaluated effects of serial passage on the biological characteristics of human umbilical cord Wharton's jelly-derived MSCs (WJ MSCs). Methods. Biological properties of WJ MSCs in the early (less than 10 passages, P10), middle (P11–20), and late (more than P20) phases including cell proliferation, cell cycle, phenotype, senescence, oncogene expression, stemness marker expression, and differentiation capacity were evaluated using flow cytometry, real-time PCR, immunocytofluorescence, and western blot. Results. It was found that there were no significant differences in cell proliferation, cell cycle, phenotype, and stemness marker expression in different phases. However, the expression of senescence-related gene, p21, and oncogene, c-Myc, was significantly upregulated in the late phase, which had close relations with the obviously increased cell senescence. Moreover, cardiac differentiation capability of WJ MSCs decreased whereas the propensity for neural differentiation increased significantly in the middle phase. Conclusions. This study reveals that WJ MSCs in the early and middle phases are relatively stable, and effect of serial passage on the lineage-specific differentiation should be considered carefully.

Gelatin hydrogel has great potential in regenerative medicine. The degradation of gelatin hydrogel is important to control the release profile of encapsulated biomolecules and regulate in vivo tissue repair process. As a plasticizer, PEG can significantly improve the mechanical property of gelatin hydrogel. However, how preparation parameters affect the degradation rate of gelatin-PEG composite hydrogel is still not clear. In this study, the significant effect factor, glutaraldehyde (GA) concentration, was confirmed by means of Plackett-Burman method. Then a mathematical model was built to predict the degradation rate of composite hydrogels under different preparation conditions using the response surface method (RSM), which was helpful to prepare the certain composite hydrogel with desired degradation rate. In addition, it was found that gelatin-PEG composite hydrogel surface well supported the adhesion and growth of human mesenchymal stem cells (MSCs). Moreover, PEG concentration not only could adjust hydrogel degradation more subtly, but also might increase the cross-linking degree and affect the cell migration. Therefore, these results would be useful to optimize the preparation of gelatin-PEG composite hydrogel for drug delivery or tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3149-3156, 2016.

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The gelatin film has great potential in biomedical applications, especially in wound healing. The combination of gelatin films and stem cells could further accelerate the skin regeneration. Although polysaccharide modification can improve the mechanical property and biological activity of gelatin films, information about the stability and cellular affinity is still limited. This study investigated the influence of polysaccharides on the stability and cellular affinity of gelatin films. Two kinds of gelatin‐polysaccharide composite films, including gelatin‐hyaluronic acid (G‐HA) and gelatin‐chitosan (G‐CS), were prepared in this study. It was found that G‐HA composite film had better short‐term and long‐term stability compared with G‐CS composite film. And G‐HA composite film also had better biological safety than G‐CS film. Moreover, the surface of G‐HA composite film supported the adhesion and growth of human umbilical cord Wharton's jelly‐derived mesenchymal stem cells (WJ MSCs) better than G‐CS film surface. These data illustrated that G‐HA composite film has better stability and cellular affinity compared with G‐CS film, which could be considered a promising delivery system of stem cells for further in vivo studies. Therefore, this work would be very helpful to optimize the preparation of gelatin‐polysaccharide composite films.