The present study was conducted to develop an effective method for establishment of porcine parthenogenetic embryonic stem cells (ppESCs) from parthenogenetically activated oocyte-derived blastocysts. The addition of 10% fetal bovine serum (FBS) to the medium on the 3rd day of oocyte culturing improved the development of blastocysts, attachment of inner cell masses (ICMs) onto feeder cells, and formation of primitive ppESC colonies. ICM attachment was further enhanced by basic fibroblast growth factor, stem cell factor, and leukemia inhibitory factor. From these attached ICMs, seven ppESC lines were established. ppESC pluripotency was verified by strong enzymatic alkaline phosphatase activity and the expression of pluripotent markers OCT3/4, Nanog, and SSEA4. Moreover, the ppESCs were induced to form an embryoid body and teratoma. Differentiation into three germ layers (ectoderm, mesoderm, and endoderm) was confirmed by the expression of specific markers for the layers and histological analysis. In conclusion, data from the present study suggested that our modified culture conditions using FBS and cytokines are highly useful for improving the generation of pluripotent ppESCs.
Animals ; Blastocyst/*cytology ; Cell Culture Techniques/*veterinary ; *Cell Differentiation ; Cytokines/metabolism ; Embryonic Stem Cells/*cytology ; Parthenogenesis ; Pluripotent Stem Cells/*cytology ; Swine/*physiology

BACKGROUND: Thrombocytosis can result in life-threatening thrombotic or hemorrhagic events. Anagrelide acts exclusively on megakaryocytes and has been reported as an useful agent in controlling thromobocytosis associated with chronic myeloproliferative disorders. METHODS: Seven patients with essential thrombocythemia and three with chronic myelogenous leukemia were enrolled and early responses and adverse effects of anagrelide were retrospectively analyzed. The drug was started with a dose of 2 mg/day with increases of 0.5 mg/day every 5~7 days as needed. RESULTS: Anagrelide in starting doses of 2 mg/day reduced the platelet count by 50%, or to less than 600,000/mm3, for at least 28 days in 7 of the 9 (78%) evaluable patients. Adverse effects of the drug were observed in 5 patients and generally well tolerated; headache in 4, gastrointestinal troubles in 2, palpitations and chest tightness in 1, and tinnitus in 1. Changes in hemoglobin or white blood cell counts in peripheral blood were minimal and tolerable. CONCLUSION: The present study shows that anagrelide is a useful platelet-lowering agent in whom hydroxyurea or interferon has failed. Long-term efficacy and adverse effects of the drug remain to be determined.
Headache ; Humans ; Hydroxyurea ; Interferons ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukocyte Count ; Megakaryocytes ; Myeloproliferative Disorders ; Platelet Count ; Retrospective Studies ; Thorax ; Thrombocythemia, Essential ; Thrombocytosis* ; Tinnitus