ABSTRACT
The large-scale and cost-effective production of quality-controlled human pluripotent stem cells (hPSCs) for use in cell therapy and drug discovery would ideally require a chemically defined xenobiotic-free culture system. Towards the development of such a system, costs associated with the use of recombinant proteins as supplements in basal culture media need to be reduced. Here, we describe a growth-factor-free culture medium that uses just three chemical compounds and a lower number of recombinant proteins than used in commercially available media. We show that the culture medium supports the long-term propagation of hPSCs, as confirmed by karyotype, the expression of pluripotency markers and the capacity to differentiate into cell types derived from the three embryonic germ layers. hPSCs growing in the medium were less dependent on glycolytic pathways than cells grown in medium containing growth factors. Moreover, the medium supported the generation of induced pluripotent stem cells derived from either human dermal fibroblasts or peripheral blood mononuclear cells. Our findings should facilitate the ongoing development of a completely xeno-free, chemically defined, synthetic culture system for hPSCs.
Subject(s)
Cell Culture Techniques/methods , Pluripotent Stem Cells , Cell Differentiation , Cells, Cultured , Culture Media/chemistry , Culture Media/metabolism , Fibroblasts/cytology , Fibroblasts/physiology , Humans , Intercellular Signaling Peptides and Proteins , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/physiology , Pluripotent Stem Cells/cytology , Pluripotent Stem Cells/physiologyABSTRACT
Therapeutic effects of HL for a collagen-induced arthritis (CIA) mouse models of HL-23 composed of 95mol% l-α-dimyristoylphosphatidylcholine (DMPC) and 5mol% polyoxyethylenedodecylether (C12(EO)23) in vivo were examined. Remarkably high therapeutic effects of HL-23 for CIA mouse models were obtained on the basis of clinical assessment of arthritis. The reduction of hyperplastic synovial membrane (pannus tissue) and destruction of the cartilage and bone by HL-23 was revealed on the basis of hematoxylin and eosin (HE) and safranin O staining. Furthermore, the downregulation of inflammatory cytokines such as interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, and IL-6 for CIA mouse models treated with HL-23 were investigated. Remarkably high therapeutic effects without joint swelling were obtained in CIA mouse models treated with HL-23.
Subject(s)
Arthritis, Experimental/drug therapy , Arthritis, Experimental/immunology , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/immunology , Cytokines/metabolism , Inflammation Mediators/metabolism , Liposomes/chemistry , Liposomes/therapeutic use , Animals , Arthritis, Experimental/pathology , Arthritis, Rheumatoid/pathology , Dimyristoylphosphatidylcholine/therapeutic use , Down-Regulation/drug effects , Female , Humans , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Mice , Mice, Inbred DBA , Polyethylene Glycols/therapeutic use , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Hybrid liposomes (HLs) can be prepared by simply sonicating a mixture of vesicular and micellar molecules in buffer solutions. This study aims to demonstrate inhibitory effects of HLs on the growth of fibroblast-like synoviocytes along with apoptosis and therapeutic effects of HLs in a mouse model with rheumatoid arthritis (RA). HLs composed of 95 mol% L-α-dimyristoylphosphatidylcholine (DMPC) and 5 mol% polyoxyethylene(23)dodecyl ether (C12(EO)23) were prepared by the sonication method. The inhibitory effects of HLs on the growth of human fibroblast-like synoviocytes-RA (HFLS-RA) cells in vitro and their inhibitory mechanism were examined. High inhibitory effects of HLs on the growth of HFLS-RA cells were observed. The induction of apoptosis by HLs was revealed on the basis of flow cytometric analysis. Furthermore, therapeutic effects of HLs in the mouse model with RA were examined in vivo. Our results demonstrate that HLs showed inhibitory effects on the growth of HFLS-RA cells in vitro along with apoptosis and therapeutic effects in mouse models of RA in vivo.
Subject(s)
Apoptosis/drug effects , Arthritis, Experimental , Arthritis, Rheumatoid , Cell Proliferation/drug effects , Dimyristoylphosphatidylcholine/pharmacology , Fibroblasts/drug effects , Liposomes/pharmacology , Polyethylene Glycols/pharmacology , Synovial Membrane/drug effects , Animals , Cell Line , Disease Models, Animal , Humans , In Vitro Techniques , Mice , Synovial Membrane/cytologyABSTRACT
Therapeutic effects of cationic hybrid liposomes (HL) composed of 87 mol% dimyristoyl-phosphatidylcholine (DMPC), 5 mol% polyoxyethylene (21) dodecyl ether (C12(EO)21) and 8 mol% O,O'-ditetradecanoyl-N-(α-trimethyl-ammonioacetyl) diethanolamine chloride (2C14ECl) on the metastasis of human colon carcinoma (HCT116) cells were examined in vivo. Cationic HL having a hydrodynamic diameter less than 150 nm were preserved for one month. Therapeutic effects were obtained in the hepatic metastasis mouse models of HCT116 cells after the intravenous injection of cationic HL. The histological analysis indicated the induction of apoptosis in the liver section of the hepatic metastasis mouse models treated with cationic HL in vivo. Therapeutic effects of cationic HL without any drugs on the hepatic metastasis were revealed for the first time on the basis of histological analyses in vivo.
Subject(s)
Antineoplastic Agents/therapeutic use , Apoptosis , Cations/therapeutic use , Colonic Neoplasms/pathology , Liposomes/therapeutic use , Liver Neoplasms/drug therapy , Liver/drug effects , Animals , Antineoplastic Agents/chemistry , Dimyristoylphosphatidylcholine/chemistry , Ethanolamines/chemistry , HCT116 Cells , Humans , Liposomes/chemistry , Liver/pathology , Liver Neoplasms/secondary , Mice , Mice, Inbred BALB C , Polyethylene Glycols/chemistryABSTRACT
New-type three-component cationic hybrid liposomes (HLs) composed of dimyristoylphosphatidylcholine (DMPC), polyoxyethylene(21)dodecyl ether (C(12)(EO)(21)) and O,O'-ditetradecanoyl-N-(α-trimethylammonioacetyl) diethanolamine chloride (2C(14)ECl) were produced. Cationic HLs were smaller and more stable than pure DMPC liposomes. It is noteworthy that cationic HLs could remarkably inhibit the growth of human colon cancer (HCT116) cells along with apoptosis in vitro for the first time in this study.
Subject(s)
Antineoplastic Agents/administration & dosage , Dimyristoylphosphatidylcholine/administration & dosage , Ethanolamines/administration & dosage , Myristates/administration & dosage , Polyethylene Glycols/administration & dosage , Apoptosis/drug effects , Carcinoma , Cell Proliferation/drug effects , Colonic Neoplasms , Dimyristoylphosphatidylcholine/chemistry , Ethanolamines/chemistry , HCT116 Cells , Humans , Liposomes , Membrane Potential, Mitochondrial/drug effects , Myristates/chemistry , Polyethylene Glycols/chemistryABSTRACT
Therapeutic effects of hybrid liposomes (HL) composed of l-α-dimyristoylphosphatidylcholine (DMPC) and polyoxyethylene(25) dodecyl ether (C(12)(EO)(25)) on the metastasis of human colon carcinoma (HCT116) cells were examined in vivo. Remarkably high therapeutic effects were obtained in the xenograft mouse models of colorectal cancer (CRC) liver metastases after treatment with HL-25 on the basis of relative liver weight and histological analysis of the liver tissue sections of mouse models with HE staining, and TUNEL staining for detection of apoptotic cells. The survival effects of HL-25 were obtained using xenograft mouse models of CRC liver metastases. Furthermore, with regard to pharmacokinetics, the accumulation of fluorescent labeled HL-25 was observed in the liver tissue of xenograft mouse models of CRC liver metastases for 24 h after the intravenous injection of fluorescent labeled HL-25. Therapeutic effects of HL without any drugs on the liver metastasis of human CRC were revealed for the first time in vivo.
Subject(s)
Antineoplastic Agents/pharmacology , Carcinoma/drug therapy , Colorectal Neoplasms/drug therapy , Dimyristoylphosphatidylcholine/pharmacology , Liposomes/pharmacology , Liver Neoplasms/drug therapy , Polyethylene Glycols/pharmacology , Animals , Antineoplastic Agents/chemistry , Apoptosis/drug effects , Carcinoma/mortality , Carcinoma/secondary , Colorectal Neoplasms/mortality , Colorectal Neoplasms/pathology , Dimyristoylphosphatidylcholine/chemistry , Female , Fluorescent Dyes , Humans , In Situ Nick-End Labeling , Injections, Intravenous , Liposomes/chemistry , Liver Neoplasms/mortality , Liver Neoplasms/secondary , Mice , Mice, SCID , Organ Size/drug effects , Polyethylene Glycols/chemistry , Survival Rate , Xenograft Model Antitumor AssaysABSTRACT
Inhibitory effects of HL-n composed of 95 mol% L-α-dimyristoylphosphatidylcholin (DMPC) and 5 mol% polyoxyethylenedodecylether (C(12)(EO)(n), n = 21, 23, or 25) on the growth of human rheumatoid arthritis (RA) fibroblast-like synoviocytes (HFLS-RA) in vitro were examined. Remarkably high inhibitory effects of HL-n on the growth of HFLS-RA cells were obtained. The induction of apoptosis by HL-n was revealed on the basis of TUNEL method. Furthermore, the therapeutic effects of HL-23 using mouse models of arthritis were investigated. Therapeutic effects without joint swelling were obtained in mouse models of RA treated with HL.
