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排序方式: 共有271条查询结果,搜索用时 15 毫秒
1.
A computerized mechanical cell stimulator for tissue culture: Effects on skeletal muscle organogenesis 总被引:8,自引:0,他引:8
Herman H. Vandenburgh 《In vitro cellular & developmental biology. Plant》1988,24(7):609-619
Summary A tissue culture system has been developed which can mechanically stimulate cells growing on a highly elastic plastic substratum
in a 24-well cell growth chamber. The collagen-coated substratum to which the cells attach and grow in the Mechanical Cell
Stimulator (Model I) can be repetitively stretched and relaxed by stepper motor with linear accuracy of 30 μm. The activity
controlling unit is an Apple IIe computer interfaced with the cell growth chamber via optical data links and is capable of
simulating many of the mechanical activity patterns that cells are subjected to in vivo. Primary avian skeletal myoblasts
proliferate and fuse into multinucleated myotubes in this set-up in a manner similar to normal tissue culture dishes. Under
static culture conditions, the muscle cells differentiate into networks of myotubes which show little orientation. Growing
the proliferating muscle cells on a unidirectional stretching substratum causes the developing myotubes to orient parallel
to the direction of movement. In contrast, growing the cells on a substratum undergoing continuous stretch-relaxation cycling
orients the developing myotubes perpendicular to the direction of movement. Neither type of mechanical activity significantly
affects the rate of cell proliferation of the rate of myoblast fusion into myotubes. These results indicate that during in
vivo skeletal muscle organogenesis, when substantial mechanical stresses are placed on skeletal muscle cells by both continuous
bone elongation and by spontaneous contractions, only bone elongation plays a significant role in proper fiber orientation
for subsequent functional work.
Supported by grants NS16753, AR36266, and RR05818 from the National Institutes of Health, Bethesda, MD. 相似文献
2.
Microtiter micromass cultures of limb-bud mesenchymal cells 总被引:4,自引:0,他引:4
Douglas F. Paulsen Michael Solursh 《In vitro cellular & developmental biology. Plant》1988,24(2):138-147
Summary A method is described for growing high-density micromass cultures of chick and mouse limb mesenchyme cells in 96-well microtiter
plates (μTμM cultures). Rapid quantitative estimates of chondrogenic expression were obtained by automated spectrophotometric
analysis of Alcian-blue-stained cartilage matrix extracts performed in the wells in which the cells had been grown. Quantitative
estimates of myogenic expression were obtained similarly using anti-sarcomere myosin monoclonal antibody and modified ELISA
techniques. This μTμM-ELISA method may be adapted for use with other antigens for which specific antibodies are available.
These methods were used to compare cartilage and muscle differentiation in 1 to 4 d μTμM cultures grown in serum-containing
(SCM) and defined (DM) media. The DM contains minimal additives (insulin, hydrocortisone, and in some cases, ascorbate or
transferrin) and supports both chondrogenesis and myogenesis. The colorimetric analyses agree well with the morphologic appraisal
of chondrogenesis and myogenesis. Similar numbers of cartilage nodules formed in all cultures, but in DM the nodules failed
to enlarge; explaining the reduced matrix synthesis in DM as compared with SCM, and suggesting that nodule enlargement is
a discrete, serum-dependent step. Studies of selected additives to DM show that transferrin enhances myogenesis, ascorbic
acid enhances chondrogenesis, and retinoic acid inhibits chondrogenesis. Together, the μTμM system, in situ colorimetric assays
of chondrogenesis and myogenesis, and DM will allow rapid prescreening of teratogens and screening of various bioactive compounds
(e.g., hormones, growth factors, vitamins, adhesion factors) for effects on limb mesenchymal cell differentiation.
This work was supported by grants RR08006-13 (DFP) and HD05505 and HD18577 (MS) from the National Institutes of Health, Bethesda,
MD. MF-20 hybridoma supernatant was obtained from the Developmental Studies Hybridoma Bank, Department of Biology, University
of Iowa, Iowa City, Iowa 52242 (maintained by NIH grant NO1-HD62915). 相似文献
3.
Ichiro Kimura Yae Gotoh Eijiro Ozawa 《In vitro cellular & developmental biology. Plant》1989,25(3):236-242
Summary A mitogenic factor which promotes quail myoblast proliferation has been purified some 105-fold from chick embryo extract by a combination of cation-exchange chromatography and heparin-affinity chromatography. The
factor is eluted from heparin-Sepharose with 2M NaCl and is a single-chain polypeptide with an apparent molecular weight of 15000 to 17000. It is active at subnanogram level
in triggering the proliferation and thereby delaying temporarily fusion of myoblasts. It also stimulates the proliferation
of quail fibroblasts in a similar effective concentration range. For both myoblasts and fibroblasts the dose-response to the
factor is quantitatively and qualitatively comparable with that of bovine pituitary fibroblast growth factor. These observations
strongly suggest that the factor very probably corresponds to chicken fibroblast growth factor or to a closely related molecule(s)
and that it is possibly involved in the regulation of myogenesis.
This work was partly supported by a grant from the National Center of Neurology and Psychiatry (NCNP grant 86-01) of the Ministry
of Health and Welfare, Japan. 相似文献
4.
Walter Malorni Pietro L. Indovina Giuseppe Arancia Stefania Meschini Maria T. Santini 《In vitro cellular & developmental biology. Plant》1990,26(4):399-410
Summary This paper describes the microscopic evidence supporting a cesium-induced delay in the fusion of chick embryo myoblast membranes
during in vitro myogenic differentiation. We have recently demonstrated that the sharp decrease in the conductivity and permittivity
of the membranes of these myogenic cells at the time of fusion is delayed 30 h by the addition of cesium to the culture medium
(Santini et al., Biochim. Biophys. Acta 945:56–64; 1988). We report here that this delay in fusion is substantiated by direct
microscopic observation and that cesium also induces ultrastructural changes in the myoblast cells themselves. Possible mechanisms
by which cesium may cause both the delay in fusion as well as the ultrastructural changes observed are discussed.
This investigation was partially supported by an Italian Consiglio Nazionale delle Ricerche grant 85.00.304.02 (to P. L. I.). 相似文献
5.
We analyzed the fiber-type composition of the soleus muscle in rats and mice to determine whether the adult proportion of fiber types is fixed soon after birth or whether it changes during postnatal maturation. We examined muscles from animals varying in age from 1 week to 1 year using monoclonal antibodies that distinguish between fast and slow isoforms of myosin heavy chains. In cross sections of unfixed muscle containing profiles of all myofibers in the muscle, we counted the fibers that stained with antibodies to fast myosin, and in adjacent sections, those that stained positive with an antibody to slow myosin. We also counted the total number of fibers in each section. Rat soleus contained about 2500 myofibers, and mouse about 1000 at all ages studied, suggesting that myogenesis ceases in soleus by 1 week after birth or sooner. In mouse soleus, the relative proportions of fibers staining positive with fast and slow myosin antibodies were similar at all ages studied, about 60%–70% being fast and 30%–40% slow. In rat soleus, however, the proportions of fast antibody-positive and slow antibody-positive fibers changed dramatically during postnatal maturation. At 1 week after birth, about 50% of rat soleus fibers stained with fast myosin antibodies, whereas between 1 and 2 months this value fell to about 10%. In mouse, about 10% of fibers at 1 week, but none at 1 year, reacted with both fast and slow antibodies, whereas in rat, fewer than 3% bound both antibodies to a significant degree at 1 week. It is puzzling why, in rat soleus, the majority of apparently fast fibers present at 1 week is converted to a slow phenotype, whereas in mouse soleus the predominant change appears to be the suppression of fast myosin expression in a subset of fibers that expresses both myosin types at 1 week. It is possible that this may be related to differences in size and the amount of body growth between these two species. 相似文献
6.
Ines Gerson Robert L. Seecof Raymond L. Teplitz 《In vitro cellular & developmental biology. Plant》1976,12(9):615-622
Summary Cultures of embryonicDrosophila melanogaster cells were examined by electron microscopy and events in myogenesis were recorded. Thick and thin myofilaments, T-tubules
and sarcoplasmic reticulum all appeared at about the same time, 10.5 hr. This was about 5 hr after the final division of myoblasts
and about the time that muscle cells were elongating, aligning and fusing. Sarcoplasm typical of insect muscle was detected
by 18.5 hr, as were myotendonal and tendocuticular junctions. Two populations of myocytes were detected, the cytoplasm of
one more electron-dense than the other. The only previous report of myofibrilogenesis in invertebrate embryos had described
novel mechanisms. InDrosophila embryonic material, however, the sequence of myofibrilogenesis resembled that in post-embryonic insect or vertebrate material.
Mrs. Pilar Toribio-Fiorio provided excellent technical assistance, and Patricia Minter, the secretarial expertise.
This investigation was supported, in part, by NIH Grant NS9330 and the James Douglas Research Fund to Robert L. Seecof and
NIH Grant No. 1 RO1 CA17223-01 to Raymond L. Teplitz. 相似文献
7.
8.
Jon Sin Allen M. Andres David J. R. Taylor Thomas Weston Yoshimi Hiraumi Aleksandr Stotland 《Autophagy》2016,12(2):369-380
Myogenesis is a crucial process governing skeletal muscle development and homeostasis. Differentiation of primitive myoblasts into mature myotubes requires a metabolic switch to support the increased energetic demand of contractile muscle. Skeletal myoblasts specifically shift from a highly glycolytic state to relying predominantly on oxidative phosphorylation (OXPHOS) upon differentiation. We have found that this phenomenon requires dramatic remodeling of the mitochondrial network involving both mitochondrial clearance and biogenesis. During early myogenic differentiation, autophagy is robustly upregulated and this coincides with DNM1L/DRP1 (dynamin 1-like)-mediated fragmentation and subsequent removal of mitochondria via SQSTM1 (sequestosome 1)-mediated mitophagy. Mitochondria are then repopulated via PPARGC1A/PGC-1α (peroxisome proliferator-activated receptor gamma, coactivator 1 alpha)-mediated biogenesis. Mitochondrial fusion protein OPA1 (optic atrophy 1 [autosomal dominant]) is then briskly upregulated, resulting in the reformation of mitochondrial networks. The final product is a myotube replete with new mitochondria. Respirometry reveals that the constituents of these newly established mitochondrial networks are better primed for OXPHOS and are more tightly coupled than those in myoblasts. Additionally, we have found that suppressing autophagy with various inhibitors during differentiation interferes with myogenic differentiation. Together these data highlight the integral role of autophagy and mitophagy in myogenic differentiation. 相似文献
9.
Yi‐zhou Tan Dong‐dong Fei Xiao‐ning He Ji‐min Dai Rong‐chen Xu Xin‐yue Xu Jun‐jie Wu Bei Li 《Cell proliferation》2019,52(4)
L‐type voltage‐gated calcium ion channels (L‐VGCCs) have been demonstrated to be the mediator of several significant intracellular activities in excitable cells, such as neurons, chromaffin cells and myocytes. Recently, an increasing number of studies have investigated the function of L‐VGCCs in non‐excitable cells, particularly stem cells. However, there appear to be no systematic reviews of the relationship between L‐VGCCs and stem cells, and filling this gap is prescient considering the contribution of L‐VGCCs to the proliferation and differentiation of several types of stem cells. This review will discuss the possible involvement of L‐VGCCs in stem cells, mainly focusing on osteogenesis mediated by mesenchymal stem cells (MSCs) from different tissues and neurogenesis mediated by neural stem/progenitor cells (NSCs). Additionally, advanced applications that use these channels as the target for tissue engineering, which may offer the hope of tissue regeneration in the future, will also be explored. 相似文献
10.