Biological Characteristicsof Umbilical Cord-derived Mesenchymal Stem Cells And The Differentiated Sweat Gland-like Cells Under Animal Serum-free Culture Conditions

Deep burn or trauma can cause serious damage to the normal structure and functionof the skin and skin appendages.The structure and function damage of the sweat glandswill seriously affect the lifequality of patients. There are many studies thatmesenchymal stem cells (MSCs) could participate effectively in promoting repair andregeneration of skin and sweat glandsand might improve repair effects ofpatients. MSCis a kind of self-renewal and pluripotency cells in research and clinical applications. Ourprevious study found that bone marrow-derived MSCs (Bone marrow derived-MSCs,BMMSCs) in the specific induction environment can successfully differentiate into thesweat glands like cells and repair damaged sweat glands. However, limited by cellculture cycle, it cannot be applied to burn and trauma and BMMSCs puncture wouldcause some pain to the donor. Also,proliferation and differentiation potential ofBMMSCs would declinewith increaseddonor age. Therefore, new sources of MSCs forstem cell research are necessary for sweat gland regeneration.Umbilical cord as a reliable source of MSCs, compared with the bone marrow iseasy to get, easy to operate, a rich source of differentiation ability, no harm to the donor,graft-versus-host disease(GVHD), a low incidence of stem cell proportion and avoidethical controversy, and many other advantages, more may be used in clinicaltreatment.Hence, using umbilical cord-derived MSCs (umbilical cord derived-MSCs,UCMSCs) has been becoming a new strategy of regenerative medicine including skinregeneration. AS the pluripotency has been confirmed, UCMSCs can be used as a newcell therapy strategies applied to sweat gland repair and regeneration research.Preliminary findings in our previous study showed that UCMSCs had the potential todifferentiate into sweat gland-like cellsunder the specific induction environment. However, due to thedifferent culture conditions,serumandcytokines, it may cause thedifferenceresults in different batches of the cell phenotype, proliferation ability, andlong-term cultured cell function and differentiation ability into sweat gland cells.Therefore, standards of culture conditionsin accordance with the clinical use andnon-animal-derived serum as well as standardization of differentiated induce system arenecessary to establish, in order to carry out further basic research and in-depthpre-clinical experimental basis.The study is divided into two parts as follows:1.Establishmentof animal origin freeculture and investigation of the biological features (expression of surface antigens andcell proliferation activity) of UC-MSCs expanded with animal serum free culture media(ASFCM).2. Establishment of animal origin freeinduction systemand determine thecapability of differentiating into sweat gland-like cells using RT-PCR, Western-blot andkaryotype analysis. Experimental results show that UCMSCs in animal serum cultureconditions are still able to maintain the biological phenotype and proliferative capacityof MSCs, and no significant difference was found between the groups. By inducedbyanimal serum sweat gland, UCMSCs can differentiateinto the sweat glands like cell andpositively expressed sweat glands markers CEA, CK14, CK19and sweat glandsdevelopmental genes EDA and EDAR.Taken together, UCMSCs can maintain thebiological features and capabilityofdifferentiatedsweat gland-like cells under safelyASFCM for clinical purposes, which provide the scientific and theoretical basis forsweat gland repair in clinical therapy and for the future establishment of sweat glandcell bank.