Selection of common markers for bone marrow stromal cells from various bones using real-time RT-PCR: Effects of passage number and donor age
TISSUE ENGINEERING
Authors: Igarashi, Akira; Segoshi, Kazumi; Sakai, Yuhiro; Pan, Haiou; Kanawa, Masami; Higashi, Yukihito; Sugiyama, Masaru; Nakamura, Kozo; Kurihara, Hidemi; Yamaguchi, Satoru; Tsuji, Koichiro; Kawamoto, Takeshi; Kato, Yukio
Abstract
Bone marrow stromal cells (BMSCs) are valuable in tissue engineering and cell therapy, but the quality of the cells is critical for the efficacy of therapy. To test the quality and identity of transplantable cells, we identified the molecular markers that were expressed at higher levels in BMSCs than in fibroblasts. Using numerous BMSC lines from tibia, femur, ilium, and jaw, together with skin and gum fibroblasts, we compared the gene expression profiles of these cells using DNA microarrays and low-density array cards. The differentiation potential of tibia and femur BMSCs was similar to that of iliac BMSCs, and different from jaw BMSCs, but all BMSC lines had many common markers that were expressed at much higher levels in BMSCs than in fibroblasts; several BMSC markers showed discrete expression patterns between jaw and other BMSCs. The common markers are probably useful in routine tests, but their efficacy may depend upon the passage number or donor age. In our study the passage number markedly altered the expression levels of several markers, while donor age had little effect on them. Considering the effects of in vivo location of BMSCs and passage, magnitude of increase in expression levels, and interindividual differences, we identified several reliable markers - LIF, IGF1, PRG1, MGP, BMP4, CTGF, KCTD12, IGFBP7, TRIB2, and DYNC1I1 - among many candidates. This marker set may be useful in a routine test for BMSCs in tissue engineering and cell therapy.
Behavioural endophenotypes in mice lacking the auxiliary GABA(B) receptor subunit KCTD16
BEHAVIOURAL BRAIN RESEARCH
Authors: Cathomas, Flurin; Sigrist, Hannes; Schmid, Luca; Seifritz, Erich; Gassmann, Martin; Bettler, Bernhard; Pryce, Christopher R.
Abstract
Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the brain and is implicated in the pathophysiology of a number of neuropsychiatric disorders. The GABA(B) receptors are G-protein coupled receptors consisting of principle subunits and auxiliary potassium channel tetramerization domain (KCTD) subunits. The KCTD subunits 8, 12, 12b and 16 are cytosolic proteins that determine the kinetics of the GABA(B) receptor response. Previously, we demonstrated that Kctd12 null mutant mice (Kctd12(-/-)) exhibit increased auditory fear learning and that Kctd12(+/-) mice show altered circadian activity, as well as increased intrinsic excitability in hippocampal pyramidal neurons. KCTD16 has been demonstrated to influence neuronal excitability by regulating GABA(B) receptor-mediated gating of post synaptic ion channels. In the present study we investigated for behavioural endophenotypes in Kctd16(-/-) and Kctd16(+/-) mice. Compared with wild-type (WT) littermates, auditory and contextual fear conditioning were normal in both Kctd16(-/-) and Kctd16(+/-) mice. When fear memory was tested on the following day, Kctd16(-/-) mice exhibited less extinction of auditory fear memory relative to WT and Kctd16(+/-) mice, as well as more contextual fear memory relative to WT and, in particular, Kctd16(+/-) mice. Relative to WT, both Kctd16(+/-) and Kctd16(-/-) mice exhibited normal circadian activity. This study adds to the evidence that auxiliary KCTD subunits of GABA(B) receptors contribute to the regulation of behaviours that could constitute endophenotypes for hyper-reactivity to aversive stimuli in neuropsychiatric disorders. (C) 2016 Elsevier B.V. All rights reserved.