Fiber-shaped cellular constructs have attracted increasing attention in the regeneration of blood vessels, nerve networks, and skeletal myofibers. Nevertheless, the generation of functional fiber-shaped cellular constructs suffers from limited appropriate microfiber-based fabrication approaches and the maintenance of regenerated tissue functions. Herein, we demonstrate a silicone-tube-based coagulant bath free method to fabricate tens of centimeters long cell-laden microfibers using single UV exposure without pretreatment of nozzles or microchannels. By modulating the exposure time, the gelatin methacrylate microfibers with tissue-like microstructures and mechanical properties are obtained. Then, a culture system integrated with a pillar well-array based stretching device is used to apply uniaxial stretching with various strain ratios in situ to cell-laden microfibers in a 60 mm petri dish. Cells with improved spreading, elongation, and alignment are obtained under uniaxial stretching. Moreover, the promotional effects of uniaxial stretching on the differentiation of C2C12 myoblasts, the formation, and contractility of myofibers become more pronounced with increasing strain ratio and achieve saturation level as strain ratio up to ∼35%.
The mechanism of anther abortion in a male sterile (ms) line (Longyou 9S) of Brassica napus L. was evaluated by a combination of light and electron microscopies. Light microscopy showed that the tapetal cells of the ms line appeared smaller than those of its fertile line, sporogenous cells were rich in vacuoles, and pollen mother cells dismantled before the tetrad stage. Electron microscopy demonstrated that sporogenous cells were also rich in very long, plate-shaped endoplasmic reticulum cisternae that actively encircled portions of cytoplasm with organelles. In later stages, many large membrane-bound inclusions were observed in the sporogenous cells. These membrane-bound inclusions were lined by single or multiple layer(s) of the endoplasmic reticulum membranes and contained vacuoles, ribosome, plastid, mitochondria, small membrane-bound inclusion, and/or intact nuclei. In meiotic interphase and early meiosis prophase I, many vesicle aggregations and multivesicular bodies containing numerous vesicles appeared in pollen mother cells. More interestingly, some of the multivesicular bodies lay in deep cytoplasm or near cell wall, and some appeared fused with plasmalemma and released the inner vesicles out of plasmalemma. Such released vesicles gradually dispersed and later disappeared. The observations suggest that active endoplasmic reticulum-dependent autophagic programmed cell death and multivesicular body-dependent polar vesicle trafficking are probably present in the studied male sterile line.
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