• 제목/요약/키워드: feeder-free

검색결과 37건 처리시간 0.027초

Construction of 3D Culture Medium with Elastin-like Polypeptide (ELP) Hydrogel for Human Pluripotent Stem Cells

  • Lee, Jonghwan;Rhee, Ki-Jong;Jung, Donjgu
    • 대한의생명과학회지
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    • 제19권1호
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    • pp.41-47
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    • 2013
  • Pluripotent stem cells (PSCs) have lots of potential in biomedical sciences owing to its potential to differentiate into any kind of cells in the body. However, it is still a challenge to culture PSCs on a large scale for application to regenerative medicine. Herein, we introduce a synthetic polymer that enables large-scale suspension culture of human PSCs. By employing suspension culture, it became unnecessary to use conventional substrata such as mouse embryonic fibroblast (MEF) or Matrigel$^{TM}$, which are believed to be main causative sources of xenogeneic contamination in cultured human PSCs in vitro. Human PSCs were cultured in the medium in which elastin-like polypeptide (ELP) dissolved. The ELP in the medium became harden as temperature increases by transforming the medium into a semi-solid gel that supported growth of human PSCs in suspension. Gel-sol transition temperature of ELP can be adjusted by modifying the peptide sequence in which 5 amino acids, Val-Pro-Gly-Xaa-Gly, repeated sequentially. We constructed 3D suspension media having transition temperature around $33{\sim}35^{\circ}C$ using an ELP consisted of 40, 60, or 80 repeats of a monomer, which was Val-Pro-Gly-Val-Gly. Among the ELPs, ELP80 was chosen as the best ELP to support growth of human PSCs in suspension culture. This result suggests that the ELP80 can be a medium component for culturing human PSCs in large-scale.

Effects of Trace Mineral Supplementation and Source, 30 Days Post-weaning and 28 Days Post Receiving, on Performance and Health of Feeder Cattle

  • Dorton, K.L.;Engle, T.E.;Enns, R.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권10호
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    • pp.1450-1454
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    • 2006
  • Three hundred and seventy-five steers (approximately 7 mo of age and $239.0{\pm}10.4kg$) were utilized to determine the effects of trace mineral (TM) supplementation and source on performance during the on-farm backgrounding and feedlot receiving phases of beef cattle production. At their respective ranches, steers were stratified by body weight into six groups. Groups were then assigned to one of six pens and pens were randomly assigned to treatments. Treatments consisted of: 1) control (no supplemental Cu, Zn, Mn, and Co), 2) inorganic trace mineral ($CuSO_4$, $ZnSO_4$, $MnSO_4$, and $CoCO_3$), and 3) organic trace mineral (iso-amounts of organic Cu, Zn, Mn, and Co). Mineral treatments were fed in alfalfa pellets formulated to supply 360 mg of Zn, 200 mg of Mn, 125 mg of Cu, and 12.5 mg of Co per head per day from either organic or inorganic trace mineral sources. Control steers received alfalfa pellets with no additional Cu, Zn, Mn, or Co. Steers were allowed free access to harvested alfalfa-grass hay throughout the 30-d on-farm backgrounding phase. On day 30 post-weaning, steers were weighed and transported to the feedlot. Steers were blocked by treatment within ranch, stratified by initial body weight, and randomly assigned to one of 36 pens (9-12 head per pen; 12 pens per treatment). Steers remained on the same on-farm backgrounding trace mineral treatments, however, trace mineral treatments were included in the total mixed growing ration. Steers were fed a corn silage-based growing diet throughout the 28 d feedlot receiving period. There was no effect of TM supplementation on performance of steers during the on-farm backgrounding phase. By the end of the 28-d feedlot receiving phase, ADG was similar between control and trace mineral supplemented steers. Steers supplemented with organic TM had greater (p<0.05) ADG than steers supplemented with inorganic TM by the end of the 28-d feedlot receiving phase. Morbidity and mortality rates were similar across treatments.

Amino acid digestibility in plant protein sources fed to growing pigs

  • Son, Ah Reum;Park, Chan Sol;Park, Kyu Ree;Kim, Beob Gyun
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권11호
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    • pp.1745-1752
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    • 2019
  • Objective: The objective was to determine standardized ileal digestibility (SID) of amino acids (AA) in 11 plant protein sources fed to growing pigs. Methods: Eleven feed ingredients used were sesame meal, two sources of soybean meal (SBM) produced in the Republic of Korea, a source of SBM produced in India, high-protein distillers dried grains (HPDDG), perilla meal, canola meal, copra meal, corn germ meal, palm kernel expeller, and tapioca distillers dried grains (TDDG). Experimental diets were prepared to contain each test ingredient as a sole source of AA, and a nitrogen-free diet was also prepared to estimate the basal ileal endogenous losses of AA. Twelve barrows surgically fitted with T-cannulas at the distal ileum with an initial body weight of 29.0 kg (standard deviation = 3.0) were individually housed in metabolism crates equipped with a feeder and a nipple drinker. A $12{\times}9$ incomplete Latin square design was employed with 12 experimental diets, 12 animals, and 9 periods. After a 5-d adaptation period, ileal digesta were collected on d 6 and 7 in each experimental period. Results: Values for apparent ileal digestibility of most indispensable AA in three sources of SBM were greater compared with other test ingredients except HPDDG and canola meal (p<0.05). Pigs fed diets containing SBM sources had also greater SID of most indispensable AA compared with those fed diets containing other test ingredients (p<0.05) except for HPDDG and canola meal. There was no difference in the apparent ileal digestibility and SID of AA among sources of SBM. The TDDG had the least value for the SID of methionine among test ingredients (p<0.05). Conclusion: The SID of most AA in SBM, HPDDG, and canola meal were greater than those in sesame meal, perilla meal, copra meal, and TDDG.

배아줄기세표의 인슐린 분비세포로의 유도 분화에 대한 연구 (Induced Differentiation of Embryonic Stem Cells to Insulin Secreting Cells)

  • 성지혜;임천규;최혜원;이형송;신현상;전진현;윤현수;궁미경
    • Clinical and Experimental Reproductive Medicine
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    • 제31권4호
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    • pp.209-215
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    • 2004
  • Objective: Embryonic stem (ES) cells could be differentiated into the specific cell types by alternation of culture condition and modification of gene expression. This study was performed to evaluate the differentiation protocol for mouse and human ES cells to insulin secreting cells. Methods: Undifferentiated mouse (JH-I) and human (Miz-hESI) ES cells were cultured on STO feeder layer, and embryoid bodies (EBs) were formed by suspension culture. For the differentiation, EBs were cultured by sequential system with three stage protocol. The differentiating ES cells were collected and marker gene expressions were analyzed by seIni-quantitative RT-PCR in each stage. Amount of secreted insulin levels in culture media of human ES cells were measured by human insulin specific RIA kit. Results: During the differentiation process of human ES cells, GATA-4, a-fetoprotein, glucose transporter-2 and Ngn-3 expression were increased whereas OctA was decreased progressively. Insulin and albuInin mRNAs were expressed from stage IT in mouse ES cells and from stage III in human ES cells. We detected 3.0~7.9 IlU/rnl secretion of insulin from differentiated human ES cells by in vitro culture for 36 days. Conclusion: The sequential culture system could induce the differentiation of mouse and human ES cells into insulin secreting cells. This is the fIrst report of differentiation of human ES cells into insulin secreting cells by in vitro culture with serum and insulin free medium.

Development of Isolation and Cultivation Method for Outer Root Sheath Cells from Human Hair Follicle and Construction of Bioartificial Skin

  • Seo, Young-Kwon;Lee, Doo-Hoon;Shin, Youn-Ho;You, Bo-Young;Lee, Kyung-Mi;Song, Key-Yong;Seo, Seong-Jun;Whang, Sung-Joo;Kim, Young-Jin;Park, Chang-Seo;Chang, Ij-Seop;Park, Jung-Keug
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.151-157
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    • 2003
  • Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

동물복지 관련 산란계 농가 실태조사 (A Farm on Laying Hens' Welfare in Korea)

  • 전중환;이준엽;박규현;김두환;송준익
    • 한국축산시설환경학회지
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    • 제18권3호
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    • pp.151-156
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    • 2012
  • 본 조사는 산란계 케이지 사육시설에 대한 대체 사육시설 등 해결방안을 모색하기 위한 기초자료를 확보하기 위하여 평사, 방사사육 및 동물복지 사육시설을 사용하는 산란계 농가를 대상으로 사육시설에 대한 실태를 조사하였다. 사육형태는 방사사육, 평사사육 및 대체 사육시설 3가지로 구분되었으며, 축사형태는 무창축사와 개방축사 2가지로 구분할 수 있었다. 급이기는 선형과 원형이 각 37.5%와 62.5%로 조사되었으며, 급수기는 니플이 75%, 선형이 12.5%, 버켓이 12.5%로 조사되었다. 사육밀도의 경우 평균 5.6수/$m^2$로 조사한 모든 농가가 인증기준에 부합하였다. 횃대가 설치되어 있지 않은 곳이 25%에 달하였으며 횃대가 설치되어 있는 농가의 경우도 수당 15 cm 이상의 횃대가 제공되어야 하는 인증기준에 미달하는 것으로 조사되었다. 또한 전체 조사농가의 50%가 재래식 산란상을 사용하고 있으며 이로 인하여 인력수거 방식으로 집란하고 있었다.

국내 담수산 조개의 섭식활동이 호수 수질에 미치는 영향 (Comparison of Filtering Abilities of Korean Freshwater Bivalves and Their Filtering Effects on Water Quality)

  • 김호섭;최광현;박정환;신재기;황순진
    • 생태와환경
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    • 제35권2호통권98호
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    • pp.92-102
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    • 2002
  • 본 연구는 담수산 조개 종간의 여과능력과 실험적으로 조성된 인공연못에서 조개의 여과섭식이 수질에 미치는 영향을 평가하기 위해 이루어졌다. 실내 실험과 인공연못에서의 섭식실험 모두에서 조개에 의한 엽록소, 총인, 부유물질과 같은 입자성 물질의 감소가 관찰되었다. 600개체의 조개가 투입된 처리구에서, 엽록소 농도와 순 1차 생산력은 $87.3{\pm}4.5\;{\mu}g/L$$106.3{\pm}8.8\;{\mu}gC\;L^{-1}\;hr^{-1}$에서 대조구와 거의 동일한 수준인 $25.0{\pm}0.5\;{\mu}g/L$$15.6{\pm}13.3\;{\mu}gC\;L^{-1}\;hr^{-1}$까지 감소하였다(P<0.05, n = 6, ANOVA). 엽록소 농도의 감소와 동시에, 투명도는 0.48m에서 1.2m까지 향상되었고, 부유물질과총인 농도는 각각 $22.0{\pm}1.0\;mg/L$에서 $7.5{\pm}0.5\;mg/L$, $133{\pm}0.8\;{\mu}g/L$에서 $7.5{\pm}0.0\;{\mu}g/L$까지 감소하였다 (P<<0.001,$r^2$>0.71,n = 11).비록 처리구에서 SRP농도의 약간의 감소와 무기질소($NH_3-N$, $NO_2-N$)의 증가가 관찰되었다 하더라도, 대조구의 농도와 비교할 때 유의적인차이는 없었다. 식물플랑크톤에 대한 여과율과 섭식과정 중에 feces와 pseudofeces와 같은 형태로의 영양염 배출율을 고려할 때, Corbicula leana가 가장 효과적인 여과섭식자였다. 이러한 결과들은 Corbicula가 입자성물질을 조절함에 있어 중요한 역할을 수행하며, 조류가 대량 발생하는 호소의 수질관리를 위해 생물학적 수질조절자로서 적용할 수 있음을 제시한다.