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Objective To establish a cultivating method for obtaining a large number of P0 generation human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Methods The hpcMSCs were isolated from human placental chorion.After primary culturing and culturing for seven days,the culture medium,the non-adherent tissue and the douching normal saline of the primary culture were centrifuged and re-cultured twice.Cell morphology was observed by an inverted microscope.CCK-8 was used to measure the cell growth curve.Flow cytometry was used to detect cell surface markers.Adipogenic and osteogenic differentiation kits were used to assess the cell differentiation potential.Results The obtained hpcMSCs were fibroblast-like adherent cells and (25.54±3.38)×106 cells were obtained per placenta.The total yield of the primary culture,secondary culture and tertiary culture were (11.73±2.09)×106,(11.12±1.42)×106 and (2.69±0.71)×106,respectively,and the incubation time were (12.00±0.64) d,(8.87±0.63) d and (12.33±0.80) d.There was significant differences in incubation time between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the incubation time of the tertiary culture had an increasing trend (P＞0.05).The yield per culture flask of the primary culture,secondary culture and tertiary culture were (1.12±0.15) × 106,(2.10±0.16)×106 and (1.04±0.16)×106,respectively.There was significant differences in the yield per culture flask between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the yield per culture flask of the tertiary culture had a decreasing trend (P＞0.05).There was no difference among the three cultures in the growth curve and the expression of surface markers,and the osteogenic and adipogenic differentiation were all positive.Conclusions The P0 generation hpcMSCs isolated from a choriocarcinoma sample can be doubled by the three cultures compared with the primary culture,which can provide plenty stem cell source for the regenerative medicine.
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Objective To observe the proliferation and differentiation properties of rat skeletal muscle satellite cells that are dissected,cultured and purified in vitro and investigate their response to stimulation by insulin-like growth factor I(IGF-I).Methods The cells were isolated from Wistar rats by two-step enzyme digestion and purified by the Percoll density centrifugation to observe their prolif- eration and differentiation properties and draw growth curve and fusion rate curve.Then,the response of the satellite cells to stimulation by IGF-I at various concentrations was determined by using MTT meth- od.Results Plenty and purified satellite cells were obtained via these methods.Conclusion IGF- I at appropriate concentration(100 ng/ml)can promote the proliferation and differentiation of rat skele- tal muscle satellite cells.
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Objective To study the cultivation of retinal progenitor cells of different ages. Methods Retinal progenitor cells of E14 and E18 SD rats were isolated,cultivated in suspension in modified DMEM/F12 serum-free medium and then differentiation was induced in vitro.Cells were observed under phase-contrast microscopy daily and identified by immunocytochemistry,scanning electron microscopy and transmission electron microscopy. Results Cultivated in DMEM/F12 serum-free medium,retinal progenitor cells formed cell spheres.After being plated,isolated cells migrated outwards and differentiated.Scanning electron microscopy demonstrated the morphology of the cell spheres and differentiated cells.Transmission electron microscopy demonstrated that there were stem cell-like cells in cell spheres and neuron-and glia-like cells after the plating.Immunocytochemistry demonstrated that most cells in spheres expressed neural stem cell marker nestin and cell division marker BrdU.After the plating,retinal progenitor cells could be induced to differentiate into various retinal cells,including Thy1.1-positive retinal ganglion cells.The percentage of retinal ganglion cells was 16.91%?4.05% at E14 and 4.65%?1.88% at E18.The differences were statistically significant(t=15.04,P
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Objective:To investigate the biological characteristics of cultured salivary gland cells in vitro. Methods: Submandibular gland cells of rat were primarily cultured and subcultured in DMEM supplemented with ?=10% fetal bovine serum. The growth of the cells was studied by cell counting and MTT assay. The morphological characteristic was observed with microscope and transmission electron microscope.The specific protein expression was investigated by immunohistochemical technic. Results:Morpholgy of the cells was epidermoid with well developed organella and zymogen granules.The cells could subultured to 4～5 passages and kept in culture for 30～40 days.The cells of primary culture and passage 2 grew faster than those of passage 4.The cells were CK8.13, keratin and ?-SMA positive. Conclusion:Rat submandibular gland cells can be primarily cultured and subcultured in vitro.The primarily cultured and first subcultured cells may be more potential in growth.
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Objective: To study the potential of growth and mineralization of cultured osteoblasts on the indium tin oxide conductive(ITO) glass disk coated with polypyrrole film (Ppy) . Methods: Osteoblasts were seeded onto ITO glass or ITO glass coated with polypyrrole film of implant materials. The cells were stained by HE and Von Kossa, observed by light microscope and scanning electron microscope (SEM) on day 3, 7, 14 and 28 after seeding respectively. Results: The osteoblast cells attached well to both ITO surfaces and Ppy films, mineralized nodules was observed on Ppy film. Conclusion: The polypyrrole films have good bone biocompatibility and it can be used as the biomaterials for dental implant.
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0.05) among the three groups in cell proliferation in 1~10 d cultures and in total protein content in 4~7 d cultures. At 4 and 7 days, ALP activity of MG63 cells cultivated on AD-TNZS disks was significantly higher than that of cells on the other samples(P
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AIM: To study the protective effects of Dachuanxiong Decoction (Rhizoma Chuan Xiong, Rhizoma Gastrodiae, etc) on injury induced by ischemia in nerve cells. METHODS:The level variation of activity and calcium ions of nerve cell were observed, choosing nerve cell ischemia model with hypoxia device and adapting confocal laser scanning microscopy and MTT method.. RESULTS:Dachuanxiong Decoction adjusted the abnormal structure of ischemia nerve cell, inhanced the survival rate and nerve call's, activity and decreased the concentration of calcium ions of nerve cell. CONCLUSION: Dachuanxiong Decoction blocked the calcium ion channels, nourished and protected nerve cells from ischemic injury.