• 제목/요약/키워드: In Vitro Development

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Potency and plasma protein binding of drugs in vitro-a potentially misleading pair for predicting in vivo efficacious concentrations in humans

  • Yim, Dong-Seok
    • The Korean Journal of Physiology and Pharmacology
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    • 제23권4호
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    • pp.231-236
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    • 2019
  • In drug discovery or preclinical stages of development, potency parameters such as $IC_{50}$, $K_i$, or $K_d$ in vitro have been routinely used to predict the parameters of efficacious exposure (AUC, $C_{min}$, etc.) in humans. However, to our knowledge, the fundamental assumption that the potency in vitro is correlated with the efficacious concentration in vivo in humans has not been investigated extensively. Thus, the present review examined this assumption by comparing a wide range of published pharmacokinetic (PK) and potency data. If the drug potency in vitro and its in vivo effectiveness in humans are well correlated, the steady-state average unbound concentrations in humans [$C_{u_-ss.avg}=f_u{\cdot}F{\cdot}Dose/(CL{\cdot}{\tau})=f_u{\cdot}AUCss/{\tau}$] after treatment with approved dosage regimens should be higher than, or at least comparable to, the potency parameters assessed in vitro. We reviewed the ratios of $C_{u_-ss.avg}$/potency in vitro for a total of 54 drug entities (13 major therapeutic classes) using the dosage, PK, and in vitro potency reported in the published literature. For 54 drugs, the $C_{u_-ss.avg}$/in vitro potency ratios were < 1 for 38 (69%) and < 0.1 for 22 (34%) drugs. When the ratios were plotted against $f_u$ (unbound fraction), "ratio < 1" was predominant for drugs with high protein binding (90% of drugs with $f_u{\leq}5%$; i.e., 28 of 31 drugs). Thus, predicting the in vivo efficacious unbound concentrations in humans using only in vitro potency data and $f_u$ should be avoided, especially for molecules with high protein binding.

Effects of Hormones and Glucose Levels during the In Vitro Culture in Medium on In Vitro Fertilization and Developmental Rates of Porcine Oocytes (돼지 수정란의 체외수정 및 발생에 미치는 호르몬 및 Glucose 첨가의 영향에 관한 연구)

  • 김상근;이명헌
    • Journal of Embryo Transfer
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    • 제9권3호
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    • pp.235-241
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    • 1994
  • The study was conducted to determine the optimal hormone and glucose levels during the in vitro culture of bovine oocytes matured and fertilized in vitro for blastocyst development. Oocytes matured in TCM 199 + 10% FCS + hormones and glucose were fertilized in vitro in a TALP medium with swim up separated and heparin-treated epididymal cauda spermatozoa. Oocytes were cultured for 2~5 days in synthetic oviduct fluid medium (SOFM) supplemented with 10% FGS and with different hormone and glucose levels, and further cultured 5 days same medium in SOFM. The results are summarized as follows : The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + $\beta$ estradiol, PMSG + $\beta$ estradiol 0 to20 hours after insemination were 88.0% and 81.8%, 82.6% and 68.4%, 80.0% and 75.0%, 80.0% and 65.0%, 77.3% and 64.7%, respectively. The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + $\beta$ estradiol, PMSG + $\beta$ estradiol 20 to 40 hours after insemination were 92.0% and 87.0%, 92.0% and 82.6%, 91.3% and 81.0%, 85.2% and 73.9%, 87.5% and 81.0%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose lelvels 0~3 days after insemination were 31.5~48.1% and 10.0~16.7%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose levels 4~8 days after insemination were 30.0~53.8% and 8.7~19.2%, respectively. The cleavage and in vitro developmental rates to blastocyst were higher in TCM 199 media containing various glucose levels 0~3 days after insemination than 4~8 days.

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Effect of Extracellular Matrix Proteins on the In Vitro Development of Parthenogenetic Mouse Eggs (세포외 기질 단백질이 생쥐 단위발생란의 체외 발달에 미치는 영향)

  • 곽대오;김선구;김영수;박충생
    • Journal of Embryo Transfer
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    • 제8권2호
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    • pp.83-90
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    • 1993
  • To investigate the effect of extracellular matrix proteins on the in vitro development of ethanol-induced parthenogenetic eggs of ICR strain mice, those were cultured in vitro in fibronectin, gelatin, or collagen precoated culture dishes containing 1.5 ml of NaH-C03$_3$-BMOC-3 medium at 37$^{\circ}C$ for 96 hrs. under the atmosphere of 5% $CO_2$ and 95% air. Fibronectin, gelatin, or collagen significantly(P$\pm$1.4, 45.4i1.4, and 44.8$\pm$O.9, respectively. And the diameter of those eggs ranged 104.6$\pm$1.9, 102.8$\pm$2.3, and 103.4$\pm$O.8 $\mu$m, respectively.

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Effect of the Combination Hot Water - Calcium Chloride on the In Vitro Growth of Colletotrichum gloeosporioides and the Postharvest Quality of Infected Papaya

  • Ayon-Reyna, Lidia Elena;Lopez-Valenzuela, Jose Angel;Delgado-Vargas, Francisco;Lopez-Lopez, Martha Edith;Molina-Corral, Francisco Javier;Carrillo-Lopez, Armando;Vega-Garcia, Misael Odin
    • The Plant Pathology Journal
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    • 제33권6호
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    • pp.572-581
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    • 2017
  • Anthracnose of papaya fruit caused by the fungus Colletotrichum gloeosporioides is one of the most economically important postharvest diseases. Hot water immersion (HW) and calcium chloride (Ca) treatments have been used to control papaya postharvest diseases; however, the effect of the combination HW-Ca on the pathogen growth and the development of the disease in infected papaya fruit has been scarcely studied. The aim of this study was to evaluate the effect of the HW-Ca treatment on the in vitro growth of C. gloesporioides conidia and the quality of infected papaya. In vitro, the HW-Ca treated conidia showed reduced mycelial growth and germination. In vivo, the HW-Ca treatment of infected papaya delayed for 5 days the onset of the anthracnose symptoms and improved the papaya postharvest quality. The combined treatment HW-Ca was better than any of the individual treatments to inhibit the in vitro development of C. gloeosporioides and to reduce the negative effects of papaya anthracnose.

Anatomical Observation of Vitrified and Glaucous Leaf from Rehmannia glutinosa Plant Produced in Vitro (지황 기내배양시 투명화된 잎과 정상잎간의 조직학적 관찰)

  • 백기엽;유광진;박상일;신성련
    • Korean Journal of Plant Tissue Culture
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    • 제24권6호
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    • pp.323-327
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    • 1997
  • Addition of growth inhibitors such as ancymidol, ABA, chloromequat, and pachlobutrazol into MS medium had no effect to preventing vitrification in cultures of Rehmania glutinosa. Anatomical investigation revealed that vitrified thick leaf tissue in vitro had larger intercellular space with poor development of sponge and pallisade tissue compared to those of in vitro grown glaucous and field grown plants. In vitro grown glaucous leaf had smaller and round type stomata showing distinguishable guard and subsidiary cell than those of reestablished plantlets into soil whereas abnormal stomata and poor development of epicuticular wax on the surface of leaf was observed in verified plantlet.

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Effect of Pantothenic Acid, Myo-Inositol, and Folic Acid on In Vitro Development of Parthenogenetic Pig Embryos (Pantothenic Acid, Myo-Inositol 및 Folic Acid가 돼지 단위 발생 배아의 체외발육에 미치는 영향)

  • You, Jin-Young;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • 제25권1호
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    • pp.1-7
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    • 2010
  • The objective of this study was to examine the effect of vitamin B (pantothenic acid, folic acid, and myo-inositol) that was supplemented to embryo culture medium on in vitro development of parthenogenetically activated (PA) pig embryos. Cumulus-oocyte complexes derived from slaughtered ovaries were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (hCG and eCG) for the first 22 h and then further cultured in hormone-free medium for an additional 22 h. After maturation culture, metaphase II oocytes that extruded 1st polar body were electrically activated and treated with $5.0\;{\mu}g/ml$ cytochalasin B for 4 h. Then, PA embryos were cultured for 7 days in a modified NCSU-23 that was supplemented with pantothenic acid, myo-inositol, or folic acid at different concentrations ($3{\sim}300\;{\mu}M$) according to the experimental design. Myo-inositol added to culture medium did not show any beneficial or inhibitory effects on embryo cleavage and blastocyst formation. However, $300\;{\mu}M$ pantothenic acid significantly inhibited blastocyst formation compared to control (no addition) (24% vs. 36%, p<0.05). Folic acid ($300\;{\mu}M$) significantly (p<0.05) increased blastocyst formation (56%) compared to control (41%). Our results demonstrated that in vitro development of PA embryos was significantly influenced by vitamin B and addition of $300\;{\mu}M$ folic acid to culture medium improved in vitro development of pig PA embryos.

In Vitro Development of Mouse Embryos in Culture Supernatant of Bovine Oviductal Epithelial Cell (소 난관 상피세포의 배양 상층액에서 생쥐 배의 체외발달)

  • 김선구
    • Korean Journal of Animal Reproduction
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    • 제22권2호
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    • pp.111-117
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    • 1998
  • This study was conducted to examine the effect of culture supernatant of bovine oviductal epithelial cell(BOEC) on in vitro development of mouse embryos. To obtain the culture supernatant, ampullary epithelial cell, ithmic epithelial cell and ciliated eptithelial cell of bovine oviduct were cultured in Ham's F-10 su, pp.emented with 10% FCS. The development rates of mouse embryos to blastocyst stage were significantly(P<0.05) higher in BOEC-culture supernatant(72.3∼82.3%) than in Ham's F-10(50.7%). The proportions of embryonic development into hatched blastocysts were significantly(P<0.05) higher in ampullary cell supernatant(43.2%), ithmic cell supernatant(48.4%) and ciliated cell supernatant (27.7%) than in Ham's F-10(14.4%). On the other hand, the effect of ciliated cell supernatant was lower than those of other cell supernatants(P<0.05). And there was no difference between ampullary cell supernatant and ithmic cell supernatnat.

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