ABSTRACT
OBJECTIVE: Human embryonic stem cells (hESCs) have recently been reported as an unlimited source of mesenchymal stem cells (MSCs). The present study not only provides an identical and clinically compliant MSC source derived from hESCs (hESC-MSCs), but also describes the immunomodulative effects of hESC-MSCs in vitro and in vivo for a carbon tetrachloride (CCl(4))-induced liver inflammation model. METHODS: Undifferentiated hESCs were treated with Rho-associated kinase (ROCK) inhibitor and induced to fibroblast-looking cells. These cells were tested for their surface markers and multilineage differentiation capability. Further more, we analyzed their immune characteristics by mixed lymphocyte reactions (MLRs) and animal experiments. RESULTS: hESC-MSCs show a homogenous fibroblastic morphology that resembles bone marrow-derived MSCs (BM-MSCs). The cell markers and differentiation potential of hESC-MSCs are also similar to those of BM-MSCs. Unlike their original cells, hESC-MSCs possess poor immunogenicity and can survive and be engrafted into a xenogenic immunocompetent environment. CONCLUSIONS: The hESC-MSCs demonstrate strong inhibitory effects on lymphocyte proliferation in vitro and anti-inflammatory infiltration properties in vivo. This study offers information essential to the applications of hESC-MSC-based therapies and evidence for the therapeutic mechanisms of action.
Subject(s)
Embryonic Stem Cells/immunology , Immunomodulation , Mesenchymal Stem Cells/immunology , Animals , Antigens, Surface/metabolism , Base Sequence , Carbon Tetrachloride/toxicity , Cell Differentiation/immunology , Cell Line , Chemical and Drug Induced Liver Injury/immunology , Chemical and Drug Induced Liver Injury/pathology , Chemical and Drug Induced Liver Injury/therapy , DNA Primers/genetics , Embryonic Stem Cells/cytology , Embryonic Stem Cells/metabolism , Embryonic Stem Cells/transplantation , Humans , In Vitro Techniques , Lymphocyte Culture Test, Mixed , Male , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Mice , Mice, Inbred BALB C , Reverse Transcriptase Polymerase Chain Reaction , Transplantation, HeterologousABSTRACT
Bone marrow-derived mesenchymal stem cells are multipotent stem cells, an attractive resource for regenerative medicine. Accumulating evidence suggests that all-trans retinoic acid plays a key role in the development and differentiation of smooth muscle cells. In the present study, we demonstrate, for the first time, that rabbit bone marrow-derived mesenchymal stem cells differentiate into smooth muscle cells upon the treatment with all-trans retinoic acid. All-trans retinoic acid increased the expression of myocardin, caldesmon, 22-kDa smooth muscle cell-specific protein (SM22alpha), and SM-myosin heavy chains in rabbit bone marrow-derived mesenchymal stem cells, as detected by reverse transcription polymerase chain reaction (PCR). Immunostaining of SM22alpha and SM-myosin heavy chains using monoclonal antibodies also indicated smooth muscle cell differentiation of rabbit bone marrow-derived mesenchymal stem cells following the treatment with all-trans retinoic acid. In addition, more than 47% of bone marrow-derived mesenchymal stem cells demonstrated the contractile phenotype of smooth muscle cells. Western blot results showed that SM-1 and SM-2 were highly expressed in the differentiated cells. These results suggest that all-trans retinoic acid may serve as a potent agent for functional smooth muscle cell differentiation in tissue engineering.
Subject(s)
Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Multipotent Stem Cells/cytology , Multipotent Stem Cells/drug effects , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/drug effects , Tretinoin/pharmacology , Animals , Base Sequence , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Bone Marrow Cells/metabolism , Calmodulin-Binding Proteins/genetics , Calmodulin-Binding Proteins/metabolism , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , DNA Primers/genetics , Gene Expression/drug effects , In Vitro Techniques , Mesenchymal Stem Cells/metabolism , Microfilament Proteins/genetics , Microfilament Proteins/metabolism , Multipotent Stem Cells/metabolism , Muscle Proteins/genetics , Muscle Proteins/metabolism , Myocytes, Smooth Muscle/metabolism , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Rabbits , Reverse Transcriptase Polymerase Chain Reaction , Smooth Muscle Myosins/genetics , Smooth Muscle Myosins/metabolism , Trans-Activators/genetics , Trans-Activators/metabolismABSTRACT
UNLABELLED: It is suggested that nanophase hydroxyapatite (nHAP) might have advantages over conventional hydroxyapatite (cHAP) as a biomaterial for bone regeneration. To be a satisfactory candidate for bone tissue engineering, it is important to support the growth and differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). The purpose of this study is to determine whether nHAP as cell growth substrata could give better support for attachment, proliferation and osteogenic differentiation of BMSCs than cHAP. MATERIALS AND METHODS: nHAP and cHAP films were prepared as the substrata for the cell growth. BMSCs obtained from rabbit were seeded on the films. Attachment, proliferation and osteogenic differentiation of BMSCs on the two kinds of films were evaluated. RESULTS: Cell attachment ratio on nHAP films was significantly higher than that on cHAP films. Doubling time on nHAP films was significantly shorter than that on cHAP films. Amount of total proteins detected from cells cultured on nHAP films was significantly higher than that on cHAP films. However, alkaline phosphatase activity and osteocalcin content of the two groups showed no significant difference. CONCLUSIONS: nHAP films favored cell attachment and proliferation but not osteogenic differentiation of BMSCs compared with cHAP films.
