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Optimization of Human Embryonic Stem Cells into Differentiation of Dopaminergic Neurons in Vitro: II. Genetically Modified Human Embryonic Stem Cells Treated with RA/AA or b-FGF

  • 신현아;김은영;이영재;이금실;조황윤;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.75-75
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    • 2003
  • Since the establishment of embryonic stem cell, pluripotency of the cells was known to allow differentiation of the cells into various cell types consisting whole body. Several protocols have been developed to induce expression of specific genes.. However, no precise protocol that will generate a single type of the cells from stem cells has been reported. In order to produce cells suitable for transplantion into brain of PD animal model, which arouse due to a progressive degeneration of dopaminergic neurons in midbrain, human embryonic stem cell (hESC, MB03) was transfected with cDNAs cording for tyrosine hydroxylase (TH). Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by the two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA/ascorbic acid (AA), embryoid bodies (EB, for 4days) derived from hES cells were exposed to RA (10$^{-6}$ M)/AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. By indirect immunocytochemical studies, proportion of cells expressing NF200 increased rapidly from 20% at 7 days to 70 % at 28 days in RA/AA-treated group, while those cells expressing NF160 decreased from 80% at 7 days to 10% at 28 days upon differentiation in N2 medium. However, in differentiation by RA/AA treatment system, there was a significant increase in proportion of neuron maturity (73%) at day 14 after N2 medium. TH#2/MB03 cells expressing TH are >90% when matured at the absence of either bDNF or TGF-$\alpha$. These results suggested that TH#2/MB03 cells could be differentiated in vitro into mature neurons by RA/AA.

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Epidemiologic Analysis of Burns in Military Hospital

  • Choi, Jangkyu;Park, Sejin;Kim, Hyun Chul
    • Journal of Trauma and Injury
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    • v.30 no.4
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    • pp.145-157
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    • 2017
  • Purpose: We accessed epidemioloy of 908 acute burns (7 years) in the military, of injuries and propose proper educational programs to suit community. Methods: We surveyed burn demographics, circumstances of injuries, size, result of treatment. Results: The mean age was 20.6 years. The flame burns (FB) (325, 35.8%) were most common, followed scald (SB) (305, 33.6%), contact (CB) (219, 24.1%), electric (EB) (45, 5.0%) and chemical burns (ChB) (14, 1.5%). The more occurred during winter (29.7%). SB had mean 3.9% total body surface area (TBSA). The 251 (82.3%) had superficial burns by spillage of hot water/food on lower limbs (45.6%), feet (33.8%) in summer (34.8%), treated with simple dressing (92.8%). Morbidity rate was 5.6%; post traumatic stress disease (PTSD) (0.7%). FB had large wound (9.3% TBSA). The 209 (64.3%) had superficial burns by ignition to flammable oils (31.7%) and bomb powders (29.2%) on head/neck (60.3%), hands (58.6%) in summer (31.7%), autumn (30.2%). They underwent simple dressing (83.4%) and skin graft (16.0%). Morbidity rate was 18.8%; PTSD (10.5%), inhalation injuries (4.0%), corneal injury (3.7%), amputations (0.9%), and mortality rate (1.2%). CB had small (1.1% TBSA), deep burns (78.5%) by hotpack (80.4%) on lower limbs (80.4%). The more (59.8%) underwent skin graft. EB had 6.8% TBSA. The 29 (64.4%) had superficial burns by touching to high tension cable (71.1%) on hand (71.1%), upper limbs (24.4%) in autumn (46.8%). They underwent simple dressing (71.1%) and skin graft (24.4%). They showed high morbidity rate (40.0%); loss of consciousness (13.3%), nerve injuries (11.1%), neuropathy (8.9%), amputations (2.2%), and mortality rate (2.2%). Conclusions: The cook should wear apron over the boots during work. The lighter or smoking should be strictly prohibited during work with flammable liquids or bomb powders. Don't directly apply hotpack to skin for a long time. Use insulating glove during electric work. Keep to the basic can prevent severe injury and proper education is important.

Practical Applications of DNA Marker-Assisted Selection and OPU-Derived IVF Embryo Transfer for the Production of High Quality Meat in Hanwoo II. Production of IVF Embryos Derived Transvaginal Ovum Pick-up from DNA Marker-Proved Hanwoo (유전자 분석을 통하여 선발된 한우로부터 초음파 유래 체외수정란 이식에 의한 고품질 한우 생산기술의 실용화 II. DNA 검정우로부터 초음파 유래 체외수정란의 생산에 관한 연구)

  • 박희성;이지삼;진동인;박준규;홍승표;이명열;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.193-201
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    • 2001
  • This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.

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