One of the most effective and safe therapeutic methods for treating vitiligo, mixed autologous keratinocytes (KCs) and melanocytes (MCs) cultures have been used for autologous cell transplantation. However, the present transplantation method is faced with a problem that may require a large amount of skin tissue and keratinocytes have limited culture potency. We have found previously that human adipose derived stromal cells (hASCs) from aspirated fat tissue could be used in place of KCs and sufficient amounts of hASCs for transplantation could be obtained by small amount of aspirated fat tissue. The present investigation was determined the effect of ASCs on ex vivo expansion MCs for transplantation. In addition, we examined for a preservation conditions of MCs which have reported low recovery rates and a slowdown in growth after cryopreservation. Various conditions including ASCs ratio, incubation period, and additive materials for MCs cultivation was determined to improve the expansion ability of MCs. The growth rate of MCs colony was elevated 6.85 folds compared the previous conditions. These MCs showed a specific expression of immature melanocyte protein, Trp-2, but did not express the mature melanocyte proteins and markers (c-kit, CD133, and etc.) of mesenchymal stem cells that represents in ASCs feeder. Results in cryopreservation experiments were determined a preservation medium for MCs showing an increased recovery rates after thawing. The characteristics of MCs after cryopreservation using a designed medium were indicated consistent morphology and immunophenotype. In conclusion, ASCs as a feeder could be used in place of keratinocytes for ex vivo expansion of MCs. For clinical trial for vitiligo patients, efficiency experiments in preclinical state should be followed.

본 실험은 transforming growth factor-(TGF-)이 첨가된 chondrogenic induction medium(CIM)을 이용하여 인간지방조직에서 유래된 중간엽 줄기세포(human adipose tissue-derived mesenchymal stem cells, ATMSCs)의 연골형성능과 gelatin-chondroitin-glucosamine scaffold(GCG-scaffold)에 접종시킨 ATMSCs의 연골형성능을 알아

본 실험은 bioceramic을 첨가하여 만든 다공성 poly D,L-lactic-co-glycolic acid(PLGA)-scaffold가 인간 지방조직에서 유래된 중간엽 줄기세포(human adipose tissue derived mesenchymal stem cells, ATMSCs)의 골 형성과정에 효과적인지를 알아보고자 수행하였다. ATMSCs를 well plate에 접종하여 골형성 유도(osteogenic induction, OI) 배양액으로

Mesenchymal stem cells (MSCs) has been reported as multipotent progenitor cells that can be expanded rapidly in vitro and differentiated into multiple mesodermal cell type. Human MSCs have been reported to be associated with neural differentiation especially in the cholinergic phenotype in several neural system. In this study, We investigated the ability of MSCs derived human aipose tissue to differentiation into neural cells expressing Islet-1 and further differentiates into cholinergic neurons in cholinergic differentiation media. Immunocytochemistry was performed to detect the expression of Islet-1 and demonstrate characteristic of neurons and cholinergic neurons. Islet-1 was massively detected in the induction stage. Following cholinergic differentiation from Islet-1-expressing MSCs, Cholinergic neuron marker ChAT was higly expressed. Also we examined the neuroprotective effects and neural differentiation of transplanted human adipose tissue-derived mesenchymal stem cells (AT-MSCs) in ischemic stroke. For transplantation, after 3days after MCAO. animal were divided into 2 group: Group A : injected phosphate buffered saline (PBS;5 ㎕ n=10), Group B: transplanted AT-MSCs (5×105 cells, n=10). Each animal received an injection into the right penumbra region (from bregma : AP;－1.3 ㎜, ML;－4.0 ㎜, DV;－5.9 ㎜). In all animals, behavior test were performed at 1, 3, 6, 9, 12, 15 days after MCAO, that was conducted by investigators who were blined to the experimental groups. mNSS test demonstrated that motor, sensory, and balance behavior were impaired after MCAO ischemic insult. Ischemic rats that received AT-MSCs exhibited significantly improved functional performance compared with PBS injected animals and histological analysis revealed that transplanted AT-MSCs expressed marker for neuron. These results suggest that AT-MSCs can be differentiated into neuron especially in cholinergic neuron and may be a potential source of treatment for neurodegenerative disease such as stroke.

손상된 뇌신경조직내에서 신경줄기세포로부터 새로운 신경세포로의 분화가 상당히 제한되어 있어 이것이 손상된 뇌신경조직의 복구가 잘 이루어지지 않는 원인이라 여겨지고 있다. 본 연구에서는 세포배양을 통해 지방조직 중간엽 줄기세포를 도파민성 신경세포와 콜린성 신경세포로 분화를 유도하였다. 중간엽 줄기세포를 신경세포로 분화시키기 위해 N2배양액에 bFGF, EGF, dimethyl sulphoxide (DMSO)와 butylated hydroxyanisole (