• 제목/요약/키워드: culture (in vitro)

검색결과 2,910건 처리시간 0.031초

The role of sonic hedgehog signaling pathway in in vitro oocyte maturation

  • Lee, Sanghoon;Cho, Jongki
    • 한국동물생명공학회지
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    • 제36권4호
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    • pp.183-188
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    • 2021
  • In vitro maturation (IVM) of oocytes is the procedure where the immature oocytes are cultivated in a laboratory until they are mature. Since IVM oocytes generally have low developmental competence as compared to those matured in vivo, development of an optimal IVM culture system by fine-tuning culture conditions is crucial to maintain high quality. In-depth knowledge and a deep understanding of the in vivo physiology of oocyte maturation are pre-requisites to accomplish this. Within ovarian follicles, various signaling pathways that drive oocyte development and maturation regulate interaction between oocytes and surrounding somatic cells. This review discusses the sonic hedgehog (SHH) signaling pathway, which has been demonstrated to be intimately involved in folliculogenesis and oocyte maturation. Advances in elucidating the role of the SHH signaling pathway in oocyte maturation will aid attempts to improve the current inferior in vitro oocyte maturation system.

단순한정배양액 내의 Sodium Chloride 및 Macromolecules가 소 수정란의 체외발육에 미치는 영향 (Effects of Sodium Chloride and Macromolecules in Chemically Defined Culture Medium on In Vitro Development of Bovine Embryos)

  • 노상호
    • 한국수정란이식학회지
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    • 제15권2호
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    • pp.191-196
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    • 2000
  • the present study was carried out to develop a completely defined culture system and determine if high NaCl concentrations in defined (PVA added) or semi-defined (BSA added) medium is toxic to bovine embryos. Oocytes from slaughterhouse ovaries were matured and fertilized in vitro. After 30 h of insemination, only 2-cell stage embryos were selected and cultured for this experiment. The culture media used were as follows : TLP(114 mM of NaCl) + BSA (3 mg/ml), TLP + PVA (1 mg/ml), mTLP(96 mM of NaCl) + BSA, mTLP + PVA. Six to ten embryos were placed into a 30$\mu$1 drop of each medium and the embryos were examined at 10 day post-insemination without medium renewal. The experiment was replicated 4 times. All data were analyzed by chi-square. There were no significant differences among TLP-BSA, mTLP-BSA and mTLP-PVA in blastocyst development (21.6, 17.2 and 20.2%), respectively. Also, no differences were obtained in hatching rates (11.7, 9.9 and 12.2%), respecitively. However, there were significant differences between TLP-PVA (1.7% and 0.6%) and other group in blastocyst formation and hatching rates, respectively (p<0.01). Development of in vitro produced embryos cultured in BSA containing medium was not affected by high NaCl concentration, but in the completely defined medium, embryonic development was highly affected by NaCl. This study shows that reduced NaCl concentration in completely defined medium is beneficial for development of bovine pre-implantation embryos in vitro.

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체외배양에서 인간 난포액이 생쥐의 배 발달에 미치는 영향 (The Effects of Human Follicular Fluid on Embryonal Development of Mouse in In Vitro Culture)

  • 민부기;최기욱;김기석;이희섭;홍기연;이봉주;이선영;박승택
    • Clinical and Experimental Reproductive Medicine
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    • 제26권2호
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    • pp.171-178
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    • 1999
  • The follicular fluid (FF) of ovary contains various biological active products which affected on the growth of follicles and the fertilization of oocyte in physiological reproductive process of mammals. This study was designed to determine the effects of human FF on fertilization of oocyte and embryonal development in vitro culture. The FF was prepared as clear without blood contamination by needle aspiration from mature follicles of human at the time of oocytes retrieval for in vitro fertilization (IVF). As the medium for culture in vitro of embryonal cells, human tubal fluid (HTF) supplemented with follicular fluids at concentrations of 10%, 40% and pure FF were used. These effects were compared to control group of cultured embryos in HTF supplemented with 0.4% BSA (bovine serum albumin). For IVF, 64 eggs in control group, 67 eggs in 10% FF, 57 eggs in 40% FF and 64 eggs in pure FF were respectively allocated. And the rates of fertilization were almost similar in all groups as resulting 82.81% in control, 85.07% in 10% FF, 87.71% in 40% FF and 81.25% in pure FF. On the examination for embryonal cleavage from fertilized eggs, the rates of developing to 4 cell stage was similar in all groups, as results 98.11% in control, 98.27% in 10% FF and 98% in 40% FF but 78.84% in pure FF. And the rates of developing to 8-16 cell stage were significantly reduced as 44% in 40% FF and 44.23% in pure FF (p<0.05) compare to 71.69% in control media. As likewise, the rates of developing to morular stage were also significantly reduced to 36% (p<0.05) and 21.15% (p<0.01) respectively in 40% FF and pure FF. And the rates to blastocystic stage of embryo was lowest as 7.69% in pure FF (Table 1). The quality of embryonal cells on cleavage to the 8-16 cell stage was poorer, higher concentrations of FF. The rates of grade 1 in pure FF, as 23.07%, was lowest compare to those of other groups, in which the rates of grade 1 in control, 10% FF and 40% FF were 58.49%, 47.36% and 34% respectively. And on the contrary, the rate of grade 4 in pure FF was highest as 23.07%, while those were 5.66% in control, 8.77% in 10% FF and 20% in 40% FF (Table 2). On the viability of embryos, the rate of embryonal cell death was more rise, at the higher concentrations as well as longer exposure in the follicular fluid. At 48 hours after in vitro culture of embryos, the rate of survival embryos in pure FF was markedly lowered as 44.23%, compare to that of control (p<0.05). But there was not significant difference between the rates of survival embryos in each group beside the pure FF, which the rates were 77.35% in control, 70.17% in 10% FF and 60% in 40% FF respectively. And at 72 hours after in vitro culture, the rates of survival embryos were also significantly dropped to 21.15% in pure and 36% in 40% at concentration of FF compare to 62.26% in control (p<0.05, p<0.01). Finally, the rate of embryonal death at 96 hours after in vitro culture was highest as 82.69% in pure FF among all groups which those were 35.84 in control, 56.14% in 10% FF and 64% in 40% FF respectively (Fig. 1, 2, 3). In conclusion, this study suggests that the FF has no effects, in particular, to the in vitro fertilization of oocytes but exerted a bad effect to the cleavage, quality and viability of the embryonal cells during in vitro culture. However, the FF is harmful on embryonal development at conditions in higher concentration and especially on the embryos after $8{\sim}16$ cell stage.

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주목의 줄기절간 조직배양에 의한 급속 대량증식 (Rapid Micropropagation by Stem Node Culture of Japanese Yew)

  • 선정훈
    • 식물조직배양학회지
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    • 제22권6호
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    • pp.335-337
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    • 1995
  • 주목의 줄기절간 조직배양에 의한 다경줄기 유도를 위해 식물생장조절물질 중 4가지 형태의 싸이토키닌 효과를 조사 한 바, BAP, kinetin 및 thidiazuron의 경우 거의 반응을 보이지 않았으나, zeatin을 5 $\times$ $10^{-5}$M 처리하였을 경우 평균 6 개 이상의 다경줄기를 얻을 수 있었으며, 배양시료에 따라서 는 12개 이상의 다경을 형성하는 경우도 관찰되었다. 유도된 다경줄기를 지속적으로 계대배양하여 줄기를 신장시킨 후 이로부터 2 cm 크기의 기내삽목도 가능하였다. 기내발근율은 LP기본배지를 이용하는 것이 비교적 양호하였으나 효율은 3%에 지나지 않았다. 기외발근의 경우, 약 34%까지 발근율을 올릴 수 있었으며 포지에 이식시킨 조직배양묘는 1년이상 정상적인 생육을 보이고 있음이 관찰되었다.

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A review on three dimensional scaffolds for tumor engineering

  • Ceylan, Seda;Bolgen, Nimet
    • Biomaterials and Biomechanics in Bioengineering
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    • 제3권3호
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    • pp.141-155
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    • 2016
  • Two-dimensional (2D) cell culture and in vivo cancer model systems have been used to understand cancer biology and develop drug delivery systems for cancer therapy. Although cell culture and in vivo model studies have provided critical contribution about disease mechanism, these models present important problems. 2D tissue culture models lack of three dimensional (3D) structure, while animal models are expensive, time consuming, and inadequate to reflect human tumor biology. Up to the present, scaffolds and 3D matrices have been used for many different clinical applications in regenerative medicine such as heart valves, corneal implants and artificial cartilage. While tissue engineering has focused on clinical applications in regenerative medicine, scaffolds can be used in in vitro tumor models to better understand tumor relapse and metastasis. Because 3D in vitro models can partially mimic the tumor microenvironment as follows. This review focuses on different scaffold production techniques and polymer types for tumor model applications in cancer tissue engineering and reports recent studies about in vitro 3D polymeric tumor models including breast, ewing sarcoma, pancreas, oral, prostate and brain cancers.

Buffalo Rat Cell과 Platelet Derived Growth Factor가 체외수정란의 체외발육에 미치는 효과 (Effect of Buffalo Rat Liver Cell and Platelet Derived Growth Factor on the Development of In Vitro Matured/In Vitro Fertilized Bovine Oocytes)

  • 양부근;정희태;김정익
    • 한국수정란이식학회지
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    • 제10권3호
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    • pp.229-236
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    • 1995
  • The experiments reported here take advantage of the large number of in vitro matured and in vitro fertilized(IVM /IVF) bovine oocytes which can be produced, permitting the design of controlled experiments to establish a simple defined medium for the study of early embryo requirements. A total of 1,386 IVM /IVF oocytes were used to compare a simple defined medium(KSOM) with more complex culture conditions used successfully for culture of bovine embryos but do not permit study of specific requirements. All experiments were extensively replicated factorials. In Experiment 1, KSOM was superior to Menezo B$_2$ medium in producing morulae plus blastocysts from IVM /IVF oocytes(33 vs 20%, P<0.()5). The yield of morulae plus blastocysts with KSOM was 22% and with BRLC added was 30%. In Experiment 2, (a 2x2 factorial of KSOM with or without BRLC and 0, 1 ng /ml of platelet derived growth factor, PDGF) more morulae plus blastocysts (40%) were produced in KSOM-BRLC co-culture containing 1 ng /ml PDGF than in the control KSOM(12%). In Experiment 3, there was no dose response when 0, 1 and 5 ng /ml of PDGF were added. The results with simple defined KSOM medium are sufficiently promising to indicate that specific requirements of the embryo may be examined in future studies with KSOM as a base.

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Application of in vitro ovary culture for cottonwood (Populus deltoides) breeding

  • Kang, Ho-Duck;Lee, Min-Soon
    • Plant Resources
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    • 제7권2호
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    • pp.87-92
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    • 2004
  • Five different poplar hybrids were tested for rescuing embryo to elongate in vitro plantiets after hybridization. Ovaries and ovules were cultured on Woody Plant Medium (WPM) supplemented with cytokinins, 6-benzylamine (BA) and zeatin. Multiple shoots were initiated from half section of capsule with immature embryos after 21 days from pollination and tiny shoots were formed after the expansion of cotyledons in ovule cultures. Germinating response was better in intraspecific hybrids $(6.53\pm1.66)$ than interspecific crosses $(0.93\pm0.54)$ from half section of capsules on WPM medium. In general, zeatin was better than BA in inducing multiple shoots from isolated ovules. The highest average number $(19.40\pm4.53)$ of shoots was produced from immature ovules of 21 days post-pollination of WPM medium supplemented with 5.0 mg/L zeatin. The highest percentage of germination was 93% from the half section of in vitro ovary cultures. Soil acclimation was successfully conducted in cell tray containing artificially mixed soil with 96% survival rate.

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타액선 도관세포의 관류 배양 기술 개발 (Development of the Three-Dimensional Perfusion Culture Technology for the Salivary Ductal Cells)

  • 김지원;김정미;최정석
    • International journal of thyroidology
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    • 제11권2호
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    • pp.160-166
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    • 2018
  • Background and objectives: Salivary hypofunction is one of the common side effects after radioiodine therapy, and its pathophysiology is salivary ductal stenosis resulting from ductal cell injury. This study aimed to develop the functional culture environment of human parotid gland ductal cells in in vitro three-dimensional perfusion culture system. Materials and Methods: We compared plastic dish culture method and three-dimensional culture system containing Matrigel and nanofiber. Morphogenesis of reconstituted salivary structures was assessed by histomorphometry. Functional characteristics were assessed by immunohistochemistry and reverse transcription polymerase chain reaction (aquaporin 5, CK7, CK18, connexin 43, and p21). In addition, we designed the media perfusion culture system and identified higher rate of cell proliferation and expression of connexin 43 in perfusion system comparing to dish. Results: Human parotid ductal cells were well proliferated with the ductal cell characters under environment with Matrigel. In the presence of Matrigel, aquaporin 5, CK18 and connexin 43 were more expressed than 2D dish and 3D nanofiber setting. In the media perfusion culture system, ductal cells in 3D culture media showed higher cells count and connexin 43 expression compared to 2D dish. Conclusion: This in vitro ductal cell perfusion culture system using Matrigel could be used to study for radioiodine induced sialadenitis model in vivo.

체외성숙시 중.대란포의 과립막세포 첨가가 배 발달에 미치는 영향 (Effect of Addition of Granulosa Cells for Oocyte Maturation on Cleavage and Development of Bovine IVF Embryos)

  • 공일근;주영국;곽대오;노규진;박충생
    • 한국수정란이식학회지
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    • 제9권1호
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    • pp.1-6
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    • 1994
  • This experiment was investigated the effect of presence of granulosa cells from follicles of different size on bovine oocyte maturation, cleavage and development to late stage. The nuclear and cytoplasmic maturation of oocytes in the IVM-IVF system are critical for subsequent embryo development. Granulosa cells when the co-cultured with oocytes may interact with cumulus-oocytes complexes and influence the development competence of the oocytes. Granulosa cells from medium (2~6 mm) and large(>1O mm) size follicles were recovered by aspiration, washed 3 times by centrifugation at 500 x g for 5 min. and used for co-culture at a concentration of 2~3 x 106 cells/mi. The oocytes were matured in vitro (IVM) for 24 hrs. in TCM-199 supplemented with 35 $\mu$g/ml FSH, 10 $\mu$g/ml LH, 1 $\mu$g/ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro (IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro (I VC) with bovine oviductal epithelial cells for 7 to 9 days. The assessment of maturation revealed that Grade J oocytes showed significantly(P

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Comparison of Bioactive Compounds and Antioxidant Activity according to Culture Systems in Artemisia fukudo

  • Eun Bi Jang;Jong-Du Lee;Hyejin Hyeon;Yong-Hwan Jung;Weon-Jong Yoon
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2022년도 추계학술대회
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    • pp.99-99
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    • 2022
  • Artemisia fukudo is a biennial plant and has been reported to have anticancer, anti-melanogenesis, and anti-inflammatory effects. However, it is difficult to produce biomass from A. fukudo, so it is not used as a material for cosmetics or pharmaceuticals. In vitro culture can stably produce biomass throughout the year. In this study, the culture system for producing the highest biomass and bioactive substances was compared. Ex vitro plants were collected in Pyoseon-eup, Jeju island in May 2021, and in vitro culture was harvested after culturing for 8 weeks (plantlet) and 4 weeks (adventitious roots), respectively. After harvest, total polyphenol content (TPC), total flavonoid content (TFC), and DPPH scavenging activity were analyzed. In biomass production, adventitious roots (FW: 5.1 g·100 ml-1, DW: 0.6 g·100 ml-1) were about 4 times higher than that of plantlets (FW: 1.8 g·200 ml-1, DW: 0.3 g·200 ml-1). Both TPC and TFC were highest in ex vitro plants (9.2 ㎍·mL-1, 31.6 ㎍·mL-1), and were 3.0 times and 1.8 times higher than those of plantlets (3.0 ㎍·mL-1, 17.8 ㎍·mL-1), respectively. The IC50 value of DPPH scavenging activity was also the best in ex vitro plants (69.8 ㎍·mL-1), followed by root root (184.4 ㎍·mL-1) and plants (325.3 ㎍·mL-1) in that order. Through additional elicitor treatment, scale-up, and advanced compounds analysis such as HPLC, it can be used as an industrial material.

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