| Objective: To seek an efficient and rapid method for isolating and culturing human hair dermal papilla cells in vitro. To explore the possibility of cultivation of microencapsulated human hair dermal papilla cells in vitro and xenogeneic model. To assess the physical and biological properties of alginate-polylysine-alginate (APA) microcapsules and Barium-alginate (BA) beads.Methods :The scalp was transected at the dermis-hypodermis interface by an eye scissors to get subcutis with mid-to-low portions of hair follicles. Subcutaneous fat with mid-to-low portions of hair follicles was put to a small bottle and cut continuously into a mash by using an eye scissors. The mixture was digested by collagenase I at 37 C for 4-5 hours. When the fibrous sheaths became loose but still surrounding the dermal papillae, the mixture was blowed up with a pipette to help release the dermalpapillae synchronously. The mixture was transported to a centrifugal tube and mixed with supplemented Dulbecco's Modified Eagle's Medium(DMEM). Then, the mixture was settled for half a minute, the hair shafts deposited on the bottom of the centrifugal tube immediately, while the low density of free cell and dermal papillae were still suspending in the upper. Took out the upper mixture into another centrifugal tube. Repeated this step for 3-4 times. Finally, the mixture containing free cells and dermal papillae was centrifuged at 300 r.p.m. for 5 mins to separate the dermal papillae from free cells. Under a microscope, the dermal papillae were collected by a transferpette and then cultured. Adhesion of dermal papillae, cell emigration, workload and frequency of contamination were compared with that of microdissection and microdissection with digestive treatment. When human hair dermal papilla cells were successfully cultured, the cells of 4-6 passage were used for routine experimentation. Cells were enclosed in alginate-polylysine-alginate (APA) microcapsules or Barium-alginate (BA) beads respectively by using the apparatus of electrostatic microencapsulated. Then, the microencapsulated human hair dermal papilla cells were cultured in vitro and xenogeneic model. Both cells viability were investigated. Following properties were compared between alginate-polylysine-alginate (APA) microcapsules and Barium-alginate (BA) beads: (1) biocompatibility, (2) strength , (3) immunoprotectivefunction, (4) cell viability.Results: It took only 15 seconds to isolate a dermal papilla under microscope by one step digestive treatment with collagenase I, and, it reduced the contamination rate down to 0%. Compared with that of microdissection and microdissection with digestive treatment, it reduced the workload and frequence of contamination greatly. The total skill of micromanipulation that was needed was to collect the dermal papillae with a transferpette under a microscope, and it was a simple step. Not only could this very rapid and easy method reduce the workload and frequency of contamination greatly, it could remain the advantages of microdissection with digestive treatment that improve adhesion of dermal papillae and cell emigration. On the other hand, APA microcapsules possessed better biocompatibility but less strength comparing with BA beads. After microencapsulation, cells in BA beads showed higher viability than that of APA microcapsules, while the cell viability increased faster in APA microcapsules. Comparing with unmicroencapsulated cells , microencapsulated cells viability decreased. Two microcapsules could protect cells from immunologic rejection when they remained intact and smooth.Conclusions: "One step digestive treatment with collagenase I " is an efficient and rapid method to isolate and culture human hair dermal papilla cells. It is feasible for microencapsulated human hair dermalpapilla cells to be cultured in vitro and xenogeneic model. To chooseappropriate method for microencapsulation in different circumstance isnecessary.
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