| Objective: This study was designed to observe the location,morphology,and ultra-structure of telocytes in human scalp tissue.Meanwhile,to culture,sort and characterize these cells in vitro.Methods : Human scalp was used in this study.In an initial experiment,using immunohistochemical staining in combination with transmission electron microscopy to observe the location of TCs in human scalp tissue and investigate the spatial relationship between TCs and their adjacent cells.The TCs from human scalp were cultured,sorted by flow cytometry and confirmed by immunohistochemical staining with anti-CD34 and CD117 antibody.Results:With help of immunohistochemical staining,numerous CD34 positive cells were observed in the connective tissue of human skin.They were concentrated around the hair follicles,blood vessels,sweat glands,sebaceous glands and adipose lobules.The CD117 positive cells mainly located at the dermis and surrounding hair follicles and sweat glands.Under transmission electron microscopy,TCs were seen and confirmed by their special morphological features.These cells were spindle shaped with small cell bodies and thin and long processes,surrounded the stem cell cluster in the bulge region of hair follicle.The TCs from scalp were sorted by flow cytometry and culture in vitro.They exhibited typical fusiform bodies with long moniliform telopodes,and expressed positive staning with CD34 and c-kit/CD117.Conclusions:The TCs in human scalp tissue were positive for staining with CD34 and c-kit/CD117 antibodes.They surrounded the stem cells,may be involved in skin homeostasis,remodelling and regeneration.Background: Acrylate/acrylamide copolymers haveexcellent optical properties and biocompatibility and are ideal biomaterials that have been widely used in tissue engineering.Muse cells are a specific subset ofmesenchymal stem cells(MSCs)that havean excellent potential for use in regenerative medicineObjective: This study was designed to investigate the effects of acrylate/acrylamide copolymers on the adhesion,proliferation and pluripotent-like properties of Muse cells,which were derived from normal human dermal fibroblasts(NHDFs)by long-term trypsin incubation(LTT).Methods: In an initial experiment,NHDFs were digested by 0.25% trypsin.After long-term trypsin incubation,NHDFs were suspended in methylcellulose(MC)medium to form first-generationl Muse cell cluster(M-clusters)that were similar to embryonic stem cells(ESCs).The M-clusters were digested into single cells by treament with 0.25% trypsin-EDTA.Then the single Muse cells were seeded on primary microarrays containing micro-spots of 275 different mixtures of acrylate/acrylamide.Each mixture was composed of2 of 11 different monomers in various proportions,and was replicated in 4 micro-spots each.According to the adhesion and growth characteristics of Muse cells on those substrates,specific polymer candidates for Muse cells were selectedand secondary microarrayswere prepared.We then observed the effects of those specific polymer candidates on the adherence,proliferation and differentiation of Muse cells and suitable candidates for their optimal culture were identified.Results: According to the adhesion and growth patterns of Muse cells on the primary microarrays,10 suitable mixtures of acrylate/acrylamide copolymerswere identified.Muse cells grew well on6 of those combinationsand around the 4 other combinations ofthose polymer mixtures.Muse cells cultured on 3 of those combinationsproliferated and differentiated into long spindle-shaped cells that looked like fibroblasts,while Muse cells cultured on 1 combination formed clusters that were ring-shaped.Muse cells cultured on some of those combinations of acrylate/acrylamideproliferated and formedclusters that appeared to be very healthy,whereas Muse cells cultured on other combinations formed clusters that expanded outwards.Conclusions: These results identified a polymer combination that was optimum for the adhesion,proliferation and maintenance of Muse cells in an undifferentiated state. |
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