New culture medium concepts for cell transplantation.

BACKGROUND: Before cell or tissue transplantation, cells or tissues have to be maintained for a certain period in vitro using culture medium and methods. Most culture media contain substances such as pH indicators and buffers. It is not known whether some of these substances are safe for subsequent application in the transplantation of cells or tissues into the human body. We investigated culture media and methods with respect to the safety of the components in future transplantation applications. METHODS: A modified culture medium--medical fluid-based culture medium (FCM)--was designed by using various fluids and injectable drugs that are already currently permitted for use in clinical medicine. Medium components necessary for optimal cell growth were obtained from approved drugs. FCM was manufactured with adjusted final concentrations of the medium components similar to those in commercial Dulbecco's modified Eagle's medium (DMEM). In particular, 1029.40 mg/L amino acids, approximately 88.85 mg/L vitamins, 13,525.77 mg/L inorganic salts, and 4500 mg/L D-glucose comprise the high-glucose FCM. Next, human fat synovium-derived mesenchymal stem cells and rat H9c2 (2-1) cells were cultured under 2 conditions: (1) DMEM-high glucose (HG), an original commercial medium, and (2) optimized FCM-HG. We assessed the morphologies and proliferation rates of these cells. RESULTS: We observed that FCM-HG was able to induce the growth of FS-MSC and commercially available H9c2 cell. The morphologies and proliferation patterns of these cells cultured under FCM-HG showed no differences compared with cells grown in DMEM-HG. CONCLUSION: Our data suggest that FCM, which we developed for the first time according to the concept of drug repositioning, was a useful culture medium, especially in cultured cells intended for human cell transplantation.

Tranexamic acid: a "diuretic" for the peritoneal membrane?

The aim of the study was to use an animal model to investigate the mechanisms by which tranexamic acid (TNA) increases ultrafiltration (UF) volume. Thirty Sprague-Dawley rats were studied in three groups (n = 10, each group). Group 1 was dialyzed with conventional dialysis solution, group 2 with conventional fluid plus low-dose TNA (300 mg/L), and group 3 with conventional fluid plus high-dose TNA (3000 mg/L). A PD catheter was inserted into each animal, and 25 mL of the designated dialysis fluid was injected twice daily for two weeks, starting seven days after surgery. Peritoneal transport properties (PTP) were measured by modified peritoneal equilibration test (PET), and transcapillary ultrafiltration (TCUF) and lymphatic absorption (LA) were determined by using dextran 70 weekly. Histological examinations of the peritoneum were performed at the end of the experiment. Drained dialysate volume (DDV) decreased in all groups; however, a lower reduction in DDV was seen in the TNA groups. PTP increased and TCUF decreased in all groups. LA increased in Group 1, but was unchanged in the other groups. No significant histological changes of the peritoneum were observed in any group. This study confirmed that, in rats, TNA maintained UF volume by reducing LA without serious adverse effects. We may safely use TNA as a "diuretic" for the peritoneal membrane.

A subtype of adenosine receptors mediating pigment dispersion in leucophores of the medaka: evidence for an A2-receptor.

1. Adenosine and its derivatives induced dispersion of leucosomes in leucophores of the medaka, Oryzias latipes. 2. Among the purines used, 5'-N-ethylcarboxiamideadenosine was the most effective and its potency was far greater than that of adenosine, N6-L-phenylisopropyladenosine and N6-cyclohexyladenosine. 3. Methylxanthines inhibited the purine action competitively, but beta adrenergic antagonists and dipyridamole did not. 4. Beta adrenergic agonists and forskolin synergistically augmented the purine action, while Li+ blocked it competitively. 5. The results suggest that medaka leucophores possess A2 adenosine receptors on the cell membranes, the stimulation of which induces leucosome-dispersion response by increasing the cellular level of cyclic AMP through activation of adenylate cyclase activity.

Effects of forskolin, isoproterenol and lithium ions on leucophores of a teleost, Oryzias latipes: evidence for involvement of adenylate cyclase in pigment-dispersion response.

Forskolin, a stimulator of adenylate cyclase, induced a dose-dependent and reversible dispersion of pigment within fish leucophores. Li+, known as an inhibitor of the enzyme, depressed pigment-dispersion response of leucophores to either forskolin or isoproterenol, inducing an aggregation of pigment within the cells. These results indicate that Li+ acted on the cells through inhibition of forskolin- or isoproterenol-stimulated adenylate cyclase activity. The results suggest that adenylate cyclase is involved in the pigment-dispersion response of leucophores and that cAMP acts as a second messenger in the response.