Novel perspective in transplantation therapy of mesenchymal stem cells: targeting the ferroptosis pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Ex vivo culture-amplified mesenchymal stem cells (MSCs) have been studied because of their capacity for healing tissue injury. MSC transplantation is a valid approach for promoting the repair of damaged tissues and replacement of lost cells or to safeguard surviving cells, but currently the efficiency of MSC transplantation is constrained by the extensive loss of MSCs during the short post-transplantation period. Hence, strategies to increase the efficacy of MSC treatment are urgently needed. Iron overload, reactive oxygen species deposition, and decreased antioxidant capacity suppress the proliferation and regeneration of MSCs, thereby hastening cell death. Notably, oxidative stress (OS) and deficient antioxidant defense induced by iron overload can result in ferroptosis. Ferroptosis may inhibit cell survival after MSC transplantation, thereby reducing clinical efficacy. In this review, we explore the role of ferroptosis in MSC performance. Given that little research has focused on ferroptosis in transplanted MSCs, further study is urgently needed to enhance the in vivo implantation, function, and duration of MSCs.

Effects of Physical Heterogeneity Variation on Differentiation Potential of Mesenchymal Stem Cells During Senescence / 医用生物力学

With the multi-directional differentiation potential such as osteogenic differentiation, chondrogenic differentiation and adipogenic differentiation, mesenchymal stem cells (MSCs) have been widely used in basic research and clinical applications. The differentiation potential of MSCs is altered during senescence. Osteogenic differentiation potential decreases, while the lipogenic differentiation potential increases in aging MSCs. Changes in differentiation potential of MSCs during senescence are accompanied with cell physical heterogeneity variation (cell size, cell stiffness and nucleoplasmic ratio). Studies have shown that changes in physical heterogeneity of stem cells may be a key factor leading to the differences in differentiation potential of MSCs. Therefore, studies on physical heterogeneity variation of MSCs during senescence will provide a new research direction in fate prediction of stem cell. In this review, the effects of physical heterogeneity variation on differentiation potential of MSCs were summarized, and the corresponding mechanism was also discussed.

Mesenchymal stem cells: the cell replacement therapy for liver cirrhosis / 中华肝胆外科杂志

Cirrhosis is an advanced stage of liver fibrosis that causes fatal complications and and threats human health.Liver transplantation is the preferred treatment method of cirrhosis patients,but the therapeutic effect of liver transplantation is limited.In recent years,researchers at home and abroad have proposed the concept of cell replacement therapy for liver cirrhosis.Among them,mesenchymal stem cells (MSCs) have a wide range of sources,the MSCs can inhibit immunity and reduce the infiltration of inflammatory cells and release of pro-inflammatory cytokines,and they can also be induced to differentiate into hepatocytes to mitigate fibrosis,which become the preferred drug of cell replacement therapy for cirrhosis.Here,we mainly reviewed the mesenchymal stem cells from the features,the mechanisms of treatment for cirrhosis,and recent experimental studies and clinical trials.

Research progress in therapeutic action of mesenchymal stem cell in allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

Allergic rhinitis (AR) is an allergic disease of nasal mucosa, currently the consensus has been reached that there is a Th1/Th2 imbalance in AR′s pathogenesis, meanwhile there are defects in the number and/or function of Treg cells. Mesenchymal stem cells (MSCs) have characteristics of multiple differentiation potential, low immunogenicity and immunomodulation function, which can be migrated and implanted into the nasal inflammatory mucosal, recovering the Th1/Th2 immune balance and up-regulate the Treg cells by immunomodulatory function to improve AR. For the present, the treatment of AR by MSCs is basically at the animal experiment stage. This paper reviews the progress of MSCs in AR′s treatment and provides a basis for the clinical treatment of MSCs.

Effect of Nrf2 overexpression on anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells / 器官移植

Objective To investigate the effect of nuclear factor erythroid-2-related factor 2(Nrf2) on the anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells(MSCs). Methods Human embryonic kidney cells(293FT) were transfected with recombinant plasmid which overexpressed Nrf2 and helper plasmid. High-titer lentivirus which overexpressed Nrf2 were obtained. MSCs were transfected with lentivirus with Nrf2 overexpression and empty lentiviral vector to establish Nrf2-MSCs which stably overexpressed Nrf2 (Nrf2 overexpression group) and green fluorescent protein (GFP)-MSCs(control group). The expression of green fluorescent in 2 groups was observed by fluorescence microscope. The expression level of Nrf2 protein in 2 groups was measured by Western Blot. The anti-hypoxia ability of 2 groups was observed by light microscope. The anti-apoptotic ability of 2 groups was measured by flow cytometry. Results Nrf2-MSCs which stably overexpressed Nrf2 were successfully established. Western Blot analysis revealed that the expression level of Nrf2 protein in the Nrf2 overexpression group was significantly higher than that in the control group(P<0.01). After 15 h hypoxia treatment, the cell activity in the Nrf2 overexpression group was significantly higher than that in the control group. Flow cytometry showed that the apoptosis rate in the Nrf2 overexpression group was (30.9±1.4)%, significantly lower than (61.3±1.3)% in the control group(P<0.05). Conclusions Nrf2-MSCs which can stably overexpress Nrf2 possess certain anti-hypoxia and anti-apoptotic ability in hypoxia environment.

Rescue of recipient MSC functions and remission of recipient osteoporosis by donor MSC-derived exosomes through transfer of miR-26a / 实用口腔医学杂志

Objective:To explore mechanisms underlying the rescuing effects of transplanted mesenchymal stem cells (MSCs) derived exosomes on estrogen-deficient osteoporosis.Methods:Mouse estrogen-deficient osteoporosis model was constructed in 12 female C57BL6/J rats and the exosome release was regulated by siRNA.Osteogenic induction,alizarin red staining and qPCR were performed to evaluate the effects of exosomes on recipient MSC functions.The miR-26a mimics and inhibitors and qPCR were used to explore the mechanisms underlying exosome-mediated functional rescue of recipient MSCs.Results:Donor MSCs alleviated estrogen-deficient osteoporosis via exosome release,and the alleviated osteoporosis by exosomes rescued the recipient MSC functions were observed.Moreover,the rescued recipient MSC functions by exosomes transferred miR-26a were found.Conclusion:Donor MSC-derived exosomes may rescue MSC functions and may remit osteoporosis of the recipient through transfering miR-26a.

The effects of BMP9 combined with NGF on the osteogenic differentiation of C3H10T1/2 mesenchymal stem cells / 实用口腔医学杂志

Objective:To investigate the effects of BMP9 combined with NGF on the osteogenic differentiation of mesenchymal stem cells(MSCs).Methods:Recombinant BMP9 adenovirus was transfected into C3H10T1/2 cells.The cells were treated by GFP,NGF,BMP9 and BMP9 + NGF respectively.The expression level of COL1,RUNX2 was detected by RT-PCR and Western blot,ALP activity was examined by ALP kit 3,12,24,48 hours,3 and 7 days after treatment,respectively.Results:The ALP activity of BMP9 + NGF group was the highest among the 4 groups.The difference in the groups firstly appeared at 3 h after treatment.The highest expression level of RUNX2 and COL1 was detected in BMP9 + NGF group.Conclusion:NGF and BMP9 may synergisticly promote osteogenic differentiation at the early stage of osteogenic induction of C3H10T1/2 cells.

Preparation and characterization of chitosan/hyaluronic acid nanoparticles for microRNA-148b delivery / 实用口腔医学杂志

Objective:To prepare and characterize chitosan/hyaluronic acid (CS/HA) nanoparticles for miRNA-148b (miR-148b)delivery.Methods:CS/HA/miR-148b nanoparticles were prepared.The particle size,zeta potential,morphology were investigated by nano laser granulometer and TEM respectively.The encapsulation efficiency of CS/HA nanoparticles was evaluated by gel retarding analysis.The cytotoxicity and transfection efficiency of CS/HA/miR-148b nanoparticles on rat bone marrow mesenchymal stem cells (MSCs) were evaluated by CCK-8 assay,transfection assay and FCM respectively.Results:The CS/HA/miR-148b nanoparticles were discrete spherical particles with the diameter of 160 to 370 nm and zeta potential of + 15 to +41 mV.N/P ratio at 20∶1 of the nanoparticles showed the optimum encapsulation efficiency for miR-148b.CS/HA/miR-148b nanoparticles exhibited no cytotoxicity to MSCs and could deliver miR-148b to the MSCs at high transfection efficiency.Conclusion:CS/HA/miR-148b nanoparticles are safe and effective for miR-148b delivery into MSCs.

Exosome extracted from hAMSCspromotes neurological function recovery after traumatic brain injury in rats / 基础医学与临床

Objective To investigate the therapeutic effect of exosomes extracted from human adipose-derived mesenchymal stem cells(hAMSCs) on traumatic brain injury (TBI) and its possible mechanism.Methods Mesenchymal stem cells(MSCs) were isolated from healthy human adipose tissue and the exosomes were extracted by ultrafiltration.Rats were divided into four groups: sham group, PBS control group, MSCs treatment group and exosomes treatment group.24 h After TBI, the treatment group was locally injected along the lesion area, 30 μL of PBS, 2×105 MSC, 25 μg protein of exosomes respectively, the total volume was 30 μL.We performed the Modified Neurological Severity Score(mNSS) and the forelimb Foot-Fault Test in all rats before injury and at 1, 3, 7, 10, 13, 16, 21 and 30 days after TBI.The rats were sacrificed at 3 and 7 days after TBI respectively,total RNA was extracted from rat brain tissue.The expression of TNF-α and IL-1β were detected by quantitative PCR.The rats were also killed at 30 days after TBI for testing the neuronal apoptosis in lesion area by tunel-neun double imm-unofluorescence.Results Exosomes treatment significantly promotes the recovery of neurological deficits caused by TBI,and the therapeutic effect is similar to MSCs, its possible mechanism may be the inhibition of the acute inflammation and the reducing of the neurons apoptosis after TBI.Conclusions Exosomes extracted from human adipose-derived mesenchymal stem cellshas promoted neurological functionrecovery after traumatic brain injury, which will provide a new and safer TBI treatment for clinical practice.

Preclinical Study of Cell Therapy for Osteonecrosis of the Femoral Head with Allogenic Peripheral Blood-Derived Mesenchymal Stem Cells

PURPOSE: To explore the value of transplanting peripheral blood-derived mesenchymal stem cells from allogenic rabbits (rPBMSCs) to treat osteonecrosis of the femoral head (ONFH). MATERIALS AND METHODS: rPBMSCs were separated/cultured from peripheral blood after granulocyte colony-stimulating factor mobilization. Afterwards, mobilized rPBMSCs from a second passage labeled with PKH26 were transplanted into rabbit ONFH models, which were established by liquid nitrogen freezing, to observe the effect of rPBMSCs on ONFH repair. Then, the mRNA expressions of BMP-2 and PPAR-γ in the femoral head were assessed by RT-PCR. RESULTS: After mobilization, the cultured rPBMSCs expressed mesenchymal markers of CD90, CD44, CD29, and CD105, but failed to express CD45, CD14, and CD34. The colony forming efficiency of mobilized rPBMSCs ranged from 2.8 to 10.8 per million peripheral mononuclear cells. After local transplantation, survival of the engrafted cells reached at least 8 weeks. Therein, BMP-2 was up-regulated, while PPAR-γ mRNA was down-regulated. Additionally, bone density and bone trabeculae tended to increase gradually. CONCLUSION: We confirmed that local transplantation of rPBMSCs benefits ONFH treatment and that the beneficial effects are related to the up-regulation of BMP-2 expression and the down-regulation of PPAR-γ expression.

MSCs: Is This the Future Therapeutic for Cancer.

Cancer is nowadays one of the main causes of death worldwide. The numbers have shown that one in three people will develop cancer at some point in their lives. Cancer is a major issue for the whole humanity, and therefore a lot of research has been running for the past years in order to understand the mechanisms that underlie tumorigenesis and eventually cancer formation, with the perspective to discover new approaches for effective treatment. Gene therapy strategies that intended to tackle cancer systemically are often impaired by inefficient delivery of the vector to the tumor site. Several studies have shown the possibility of using mesenchymal stem cells (MSCs) as a future therapeutic mechanism against cancer, since they possess important features such as the ability to home to and target cancer cells. The engineering of MSCs to produce and deliver an apoptotic factor, calledtumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been studied excessively. TRAIL is a transmembrane protein that causes selective apoptosis of tumor cells but does not have any harmful effects on the normal neighboring cells. Experiments have shown that this approach has significant results in mouse models and has now proceeded for clinical trials.

Responses from mesenchymal stem cells to mechanical stimulation for variable duration / 医用生物力学

Objective To study mechanical responses from mesenchymal stem cells (MSCs) under different mechanical stimulus duration, by measuring its elastic modulus and characterizing its stress fibers. Methods High resolution images of MSCs cytoskeleton in vitro were acquired by using atomic force microscope (AFM) and laser scanning confocal microscope (LSCM). AFM cantilever with micro-bead attached probe was used to perform force-distance curve experiment on MSCs at the approaching time of 0.1，0.5, 1, 5,10 s, respectively. The elastic modulus of MSCs at 300 nm indentation depth were measured and compared. Results The rat MSCs cytoskeleton presented an intensely organized network structure. The elastic modulus of rat MSCs varied obviously for different mechanical stimulus duration. The median and quartile (QR) of MSCs elastic modulus were 10.02 (QR=9.66),1.94 (QR=7.71),3.63 (QR=19.33),17.15(QR=35.13), 23.52 kPa(QR=34.87), with probe approaching time at 0.1，0.5, 1, 5,10 s, respectively. The MSCs elastic modulus showed the tendency of increasing with stimulus duration increasing, except for the extremely short stimulus (0.1 s). Conclusions Unlike inorganic elastomer, rat MSCs possess complete and flexible mechanical load-bearing structure and can respond actively to a relatively longer mechanical stimulation, with an increase of elastic modulus. These results may provide basic data for further tissue engineering researches on mechanical regulation of MSCs behavior.

Effects of miR-30 a on cell cycle and dendritic cell-suppressive function of MSCs / 中国免疫学杂志

Objective:Mesenchymal stem cells( MSCs) have self-renewal capacity and potential to differentiate into the cells.It was reported that the expression of miR-30a changed in some immune diseases.But it remains unclear the effect of miR-30a on the im-munoregulatory functions of MSCs.Here we studied the impact of miR-30a on the phenotype,cell viability,apoptosis,cell cycle and im-munoregulatory functions of MSCs.Methods: The mixed enzyme methods were used for the isolation of human umbilical cord MSCs.Flow cytometry(FCM)was used to investigate the effect of overexpressed miR-30a on the phenotype of MSCs.CCK-8 was used to examine the cell viability of miR-30a-overexpressed MSCs.Annexin V/PI was used for the detection of apoptosis of MSCs.Q-PCR and Western blot were used to investigate the effect of miR-30a on the expression of Cyclin E2( CCNE2).CCNE2 was one putative target of miR-30a predicted by Targetscan database.The effects of miR-30a-overexpressed MSCs on the maturation of dendritic cells(DCs)were determined.Results:Overexpression of miR-30a blocked the cell cycle of MSCs in the G0/G1 phase by inhibiting the expression of CCNE2,but did not affect the phenotype, cell viability and apoptosis of MSCs.When co-cultured with DCs, although MSCs down-regulated the expression of CD40 and CD86 on DCs,overexpression of miR-30a more significantly enhanced the suppressive impact of MSCs on the maturation of DCs.Conclusion: miR-30a affects the cell cycle of MSCs and enhances its immunosuppressive effect on DCs.

In vitro and in vivo neurogenic potential of mesenchymal stem cells isolated from different sources.

Regenerative medicine is an evolving interdisciplinary topic of research involving numerous technological methods that utilize stem cells to repair damaged tissues. Particularly, mesenchymal stem cells (MSCs) are a great tool in regenerative medicine because of their lack of tumorogenicity, immunogenicity and ability to perform immunomodulatory as well as anti-inflammatory functions. Numerous studies have investigated the role of MSCs in tissue repair and modulation of allogeneic immune responses. MSCs derived from different sources hold unique regenerative potential as they are selfrenewing and can differentiate into chondrocytes, osteoblasts, adipocytes, cardiomyocytes, hepatocytes, endothelial and neuronal cells, among which neuronal-like cells have gained special interest. MSCs also have the ability to secrete multiple bioactive molecules capable of stimulating recovery of injured cells and inhibiting inflammation. In this review we focus on neural differentiation potential ofMSCs isolated from different sources and how certain growth factors/small molecules can be used to derive neuronal phenotypes from MSCs. We also discuss the efficacy of MSCs when transplanted in vivo and how they can generate certain neurons and lead to relief or recovery of the diseased condition. Furthermore, we have tried to evaluate the appropriatemerits of different sources ofMSCs with respect to their propensity towards neurological differentiation as well as their effectiveness in preclinical studies.

Mesenchymal stem cell-conditioned medium could ameliorated triptolide induced damage in KGN cells / 中国免疫学杂志

Objective:To explore the ameliorative effect and machanism of MSCs conditioned medium on the ovarian granulosa cells damage induced by triptolide.Methods: Cell Counting Kit 8 assay was used to examine the cell vitality of KGNs with the treatment of triptolide.The mixed enzyme digestion method were used for the isolation of human umbilical cord mesenchymal stem cells (MSCs),and flow cytometry was used for the subsequent immunotype identification.MSCs conditioned medium was collected ,and Cell Counting Kit 8 assay and PI staining was used to analyse the effect of MSCs conditioned medium on the cell vitality and cell cycle distri -bution of triptolide-damaged KGN.Real-time PCR method was used to examine the expression of cell cycle related gene CDKN1A.Results:Triptolide can inhibit KGN cell growth with the inhibition of cell vitality and cell cycle of KGN.MSCs conditioned medium did not influence the proliferation and cell cycle of normal KGN , but improved the triptolide-induced vitality inhibition and extent of S-phase arrest, and inhibit the abnormal up-regulation of CDKN1A in KGN.Conclusion: MSCs conditioned medium ameliorated the KGN cell damage induced by triptolide.

The role of PDL2 in human placenta mesenchymal stem cell-mediated inhibition of peripheral blood T cells / 中华微生物学和免疫学杂志

Objective To investigate the effect of programmed death ligand 2 (PDL2) in human placenta derived mesenchymal stem cells(hPMSCs) mediated immunoregulation on peripheral blood T cells activation,proliferation and cell cycle.Methods The expression of the PDL2 on hPMSCs was detected by RT-PCR,LSCM and FCM,respectively.Specific PDL2 siRNAs were transfected into hPMSCs via cathodolyte liposome transfection method.T cells were sorted from healthy peripheral blood by gradient centrifugation.The expression of early activation phenotype,proliferation and cell cycle of T cells were analyzed by FCM.Results PDL2 siRNA could effectively block the expression of PDL2 which was highly expressed on hPMSCs.The expression of CD69 on T cells had no significantly difference in blocking groups compared with unblocking groups.hPMSCs could inhibit the proliferation of T cells induced by PMA,compared with that of unblocking groups,the number of the T cells in G0/G1 phase was decreased while the number of the T cells in S phase was increased in the blocking groups.Conclusion PDL2 expressed on hPMSCs could promote the inhibitory effect of hPMSCs on T cell cycle and proliferation.

Role of mechanobiology in mesenchymal stem cell-based tissue engineered cartilage / 医用生物力学

Mesenchymal stem cells (MSCs) is an ideal cell type extensively used in cartilage multipotency to differentiate into cartilage and can be isolated from a wide variety of tissue sources with strong in vitro expansion. Since cartilage has the important mechanical properties, it is necessary to highlight and evaluate the mechanobiological properties of MSC-based tissue engineered cartilage. To better understand the relationship between inducing factor of cartilage repair, signal pathway and mechanical properties, this paper reviews the advances made on research of mechanobiology in MSC-based tissue engineering cartilage, discusses the existing problems in this field, and try to point out some new approaches or directions worthy of such investigation.

Isolation, culture and identification of mesenchymal stem cells from human umbilical cord as well as their osteogenic and adipogenic differentiation / 西安交通大学学报(医学版)

Obje:ctive To establish an optimized method to isolate, culture and identify human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and induce their osteogenic and adipogenic differentiation. Methods The hUCMSCs were isolated from human umbilical cord by digestion with collagenase. After serial subcultivation in vitro, the stem cells were passaged. Morphologic appearance of hUCMSCs was observed under an optical microscope and atomic force microscope. The proliferation rate was measured by MTT assay. Cell cycle and surface antigens were measured by flow cytometry. The osteogenic and adipogenic differentiation was tested and evaluated by specific staining methods. Results The isolation of hUCMSCs by digestion with collagenase was efficient. After seeded for 24 hours, the adherent cells showed spindle shape and fibroblast cell-like shape and the size of hUCMSCs was homogeneous. The similar growth curves of passage 3 and 7 exhibited a great potential for proliferation. Flow cytometry analysis revealed that CD29, CD44 and CD105 were highly expressed on the surface of passages 3 cells, but the expression was negative for CD34, CD45 and HLA-DR. After culture in inducing medium, the cells were successfully induced into osteogenic and adipogenic lineages. These cells were highly positive for alkaline phosphate staining and also showed mineralization presented with von kossa staining after 4 weeks' culture induction of osteogenic differentiation. Furthermore, liquid vacuoles were detected by oil red O staining after 3 weeks' culture induction of adipogenic differentiation. Conclusion An in vitro method for isolation and purification of hUCMSCs from human umbilical cord has been established. The cultured cells were composed of only undifferentiated cells and their biological properties were stable. The hUCMSCs are expected to be a new type of stem cells of tissue engineering.

Regulation of the Wnt Signaling Pathways during Cell Culture of Human Mesenchymal Stem Cells for Efficient Bone Regeneration / Oral Science International

Tissue engineering and bone regeneration techniques using mesenchymal stem cells (MSCs) have started to be applied to the field of oral and maxillofacial surgery. Clinically, a shortened treatment time and improved efficiency are necessary because of the patients' needs and the running cost of cell culture. In the present study, the cultivation process for human MSCs (hMSCs) was examined by regulating the Wnt signaling pathway. We activated Wnt signaling with LiCl and inhibited Wnt signaling with sFRP-3 (secreted Frizzled-Related Protein-3). The proliferation of LiCl-treated hMSCs was examined by studying the cell growth rate and performing BrdU assays. Osteogenic differentiation of sFRP-3-treated hMSCs was examined by alizarin red staining, and osteogenic gene expression on days 7 and 14 after induction was examined by reverse-transcription polymerase chain reaction (RT-PCR) analysis and quantitative real-time RT-PCR analysis. LiCl-treated hMSCs showed increased cell numbers and BrdU-positive cells as compared to the untreated cells. Alizarin red staining showed early mineralization of hMSCs on day 7 of the sFRP-3 treatment. A high expression level of the alkaline phosphatase gene on days 7 and 14 of sFRP-3 treatment was also demonstrated. These results suggest that the regulation of the Wnt signaling pathway contributes to the increased cell numbers and the early osteogenic differentiation of hMSCs. This study supports the possibility that the regulation of the Wnt signaling pathway contributes to the development of effective and efficient bone regeneration techniques.

Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells

INTRODUCTION: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. MATERIALS AND METHODS: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco's modified Eagle's medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. RESULTS: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. CONCLUSION: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.