ABSTRACT
Objective: To investigate the effects of RCCS simulated microgravity on the metabolism of the human keratinocyte cell line HaCaT. Methods: The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and HaCaT cells were cultured in vitro and divided randomly into simulated microgravity group (SMG) and normal gravity group (NG). The two group HaCaT cells were collected respectively after 1 d, 2 d and 3 d culture, and the samples were analyzed by LC/MS metabolomics. The differential metabolites between the SMG and NG cells were identified with partial least squares discriminant analysis ((O)PLS-DA), and the data were input into the KEGG database for the construction and functional analysis of metabolic pathways. Results: Comparing to NG cells, after 1 d culture, there were 74 different metabolites in SMG cells, among which 16 were up-regulated and 58 were down-regulated; after 2 d culture, there were 89 different metabolites, among which 15 were up-regulated and 74 down-regulated; after 3d culture, there were 100 different metabolites, of which 23 were up-regulated and 77 were down-regulated. The differentially expressed 49 metabolites (VIP>1 and P<0.05) after 3 d were set as target metabolites, within which the sphingosine, glutamate, and docosapentaenoic acid were down-regulated, and dehydrated sorbitol was up-regulated. KEGG analysis indicated that the metabolic pathways involved were amino acid metabolism, lipid metabolism, cell proliferation and apoptosis, substance transport, catabolism, and signal transduction. Conclusion: RCCS simulated microgravity may have significant impacts on keratinocyte metabolism mainly involving metabolites such as sphingolipids and glutamate as well as the related signaling pathways.
ABSTRACT
Objective To investigate the therapeutic effect of frozen-thawed murine islets which were transplanted into diabetic rats after cultured with hyperbaric oxygenated rotary cell culture system (HORCCS). Methods The purified rat islets were divided into two groups: A. In vitro experiment groups (IvEG) : The rat islets in each subgroup were cultured in HORCCS or common medium for 30 days, then evaluated for the intracellular DNA and insulin contents of islets, and the viability and insulin secreting level of islets. B. Islet transplantation experimental groups (TxEG) : The frozen-thawed islets were cultured in HORCCS or common medium for 7 days, and then transplanted into the recipients. We observed the blood glucose level (BGL) and insulin secreting level in the recipients as well as the uhrastructure change of islets in TxEG. Results The viability and insulin secreting level of islets cultured with HORCCS at 14th day were much higher than those cultured with common medium (P <0.05). The blood glucose level in recipients transplanted with islets cultured with HORCCS recovered to normal value at the 2nd week and lasted for 8 weeks. All these recipients maintained the normal glucose tolerance curve. Electronic microscopy found microchannel outlets on the surface of the frozen-thawed islets cultured with HORCCS. Conclusions Frozen-thawed islets cultured with HORCCS could establish nutrient transmission microchannels, which were not only capable of oxygen and nutrients transmission, but also improving cryopreservation solution to diffuse inside the islet cells evenly and uniformly. So this method not only lessens islet damage from cryopreservation, but also improves the effect of transplantation.
ABSTRACT
Objective To explore the method and technique to gain adequate seed cells for cartilage tissue engineering Methods hTERT gene was introduced into rabbit mandibular condylar chondrocytes by eukaryotic vector Rapidly proliferated immortalized chondrocytes in positive clones in micro carrier rotary cell culture system (RCCS) were screened and selected The growth of immortalized chondrocytes and the metabolic rate were observed Collagen type Ⅱ expression of immortalized chondrocytes of the experimental groups was observed The immunohistochemical results were compared with those of the control groups Results The immortalized chondrocytes in experimental groups could grow rapidly with a high metabolic rate and shorter population doubling time (PD) ( P
ABSTRACT
Objective To explore the way and technique to gain adequate seed cells for skin tissue engineering sufficiently. Methods Human telomerase reverse transcriptase (hTERT) gene was introduced into rabbit keratinocytes by eukaryotic vector. The positive clones were selected and cultured in microcarrier-RCCS. The growth of immortalized keratinocytes was observed, and the metabolic rate and pan-cytokeratins (AE1/AE3) expression of immortalized keratinocytes in experimental groups were detected and compared with those in the control group. Results The immortalized keratinocytes in the experimental groups grew rapidly and had a high metabolic rate and shorter population doubling time (PD) (P