• Title/Summary/Keyword: Colony forming units

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Ex vivo Expansion and Clonal Maintenance of CD34+ Selected Cells from Cord Blood and Peripheral Blood (제대혈 및 말포혈로부터 분리한 CD34 양성 세포의 체외 증폭 및 클론 유지)

  • Kim, Soon Ki;Ghil, Hye Yoon;Song, Sun U.;Choi, Jong Weon;Park, Sang Kyu
    • Clinical and Experimental Pediatrics
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    • v.48 no.8
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    • pp.894-900
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    • 2005
  • Purpose : Because of the unavailability of marrow transplantation, umbilical cord blood (CB) is increasingly being used. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from cord blood source and mobilized peripheral blood (PB) in a serum-free media. Methods : The CD34+ cells, selected from CB and mobilized PB, were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at culture days 0, day 4, day 7, and day 14 with various growth factors. Results : The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the PB at day 7 (2 fold increase than PB). The CB-selected CD34+ cells produced more BFU-E colonies than did the PB on culture at days 7 and at day 14. Also, the CB-selected CD34+ cells produced more CFU-Mk colonies than did the PB on culture at day 4 and at day 7. Conclusion : The ex vivo expansion of the CB cells may be promising in producing total cellular expansion, CFU-Mk and BFU-E compared with PB for 7 to 14 days. The growth factors combination including megakaryocyte growth and development, flt3-ligand and interleukin-3 showed more expansion in the view of total cells and clonal maintenance compared with less combination.

A Study on Variation of Colony Forming Units of Heterotrophic Bacteria by Input Ratios of Bulking Materials in Aerobic Composting of Food Wastes (음식물류폐기물의 호기성 퇴비화에 있어서 팽화재 투입비에 따른 타가영양세균의 균락형성단위의 변화에 관한 연구)

  • Park, Seok-Hwan
    • Journal of Environmental Health Sciences
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    • v.32 no.4 s.91
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    • pp.353-358
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    • 2006
  • This study was performed to evaluate the effects of input ratios of bulking material in aerobic composting of food wastes on variation of colony forming units of heterotrophic bacteria. Wood chips were used as a bulking material. Volume ratios of food wastes to wood chips in reactor of Control, WC-1 and WC-2 were 10/0, 10/5 and 10/10, respectively. Reactors were operated for 24 days with 1hour stirring by 1rpm and 2 hours of the forced aeration per day. WC-2 reached high temperature range faster than WC-1, and the maximum temperature of WC-2 was higher than that of WC-1. This means that the reaction velocity of composting of WC-2 was faster than that of WC-1. Judging from the profile of pH changes, composting of WC-1 proceeded slowly and continuously. Composting of WC-2 proceeded rapidly in the former half reaction period, and kept steady state of high pH in the latter half reaction period. Namely, composting of WC-2 was nearly completed in the former half reaction period. In the case of WC-1 and WC-2. the maximum temperature was followed by the rapid pH increase in 2-3 days, and this was followed by the maximum Colony Forming Units(CFU) in 3 days. But, these three items of WC-2 always appeared faster and higher than those of WC-1.

A Study on Variation of Colony Forming Units of Fungi by Input Ratios of Wood Chips in Aerobic Composting of Food Wastes (음식물류폐기물의 호기성 퇴비화에 있어서 목재세편의 투입비에 따른 곰팡이의 균락형성단위의 변화에 관한 연구)

  • Park, Seok-Hwan
    • Journal of Environmental Health Sciences
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    • v.33 no.5
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    • pp.451-455
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    • 2007
  • This study was performed to evaluate the effects of input ratios of bulking material in aerobic composting of food wastes on variation of colony forming units(CFU) of fungi. Wood chips were used as a bulking material. Volume ratios of food wastes to wood chips in reactor of Control, WC-1 and WC-2 were 10/0, 10/5 and 10/10, respectively. Reactors were operated for 24 days with I hour stirring by 1rpm and 2 hours of the forced aeration rate of $80L/min{\cdot}m^3$ per day. WC-2 reached high temperature range faster than WC-1, and the maximum temperature of WC-2 was higher than that of WC-1. WC-2 reached high pH range faster than WC-1. and the maximum pH of WC-2 was higher than that of WC-1. WC-2 reached high Log(CFU/gram) range faster than WC-I, and the maximum Log(CFU/gram) of WC-2 was higher than that of WC-1. These all mean that the reaction velocity of composting of WC-2 was faster than that of WC-1. The profile of fungi changes in Log(CFU/gram) was similar to that of temperature changes (r=0.8861) not pH changes (r=0.1631).

Enhancing effect of Canavalia gladiata DC semen on the hematopoietic expansion and function of stem cells (도두(刀豆)의 조혈증진 작용에 관한 연구)

  • Kim, Seung-Hyung;Kim, Kun-Hoae;Chi, Gyeong-Yup;Cho, In-Sik;Kim, Han-Young;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.9-16
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    • 2012
  • Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

Regular Exercise Training Increases the Number of Endothelial Progenitor Cells and Decreases Homocysteine Levels in Healthy Peripheral Blood

  • Choi, Jeong Kyu;Moon, Ki Myung;Jung, Seok Yun;Kim, Ji Yong;Choi, Sung Hyun;Kim, Da Yeon;Kang, Songhwa;Chu, Chong Woo;Kwon, Sang Mo
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.2
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    • pp.163-168
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    • 2014
  • Endothelial progenitor cells (EPCs) are known to play an important role in the repair of damaged blood vessels. We used an endothelial progenitor cell colony-forming assay (EPC-CFA) to determine whether EPC numbers could be increased in healthy individuals through regular exercise training. The number of functional EPCs obtained from human peripheral blood-derived AC133 stem cells was measured after a 28-day regular exercise training program. The number of total endothelial progenitor cell colony-forming units (EPC-CFU) was significantly increased compared to that in the control group (p=0.02, n=5). In addition, we observed a significant decrease in homocysteine levels followed by an increase in the number of EPC-CFUs (p=0.04, n=5), indicating that the 28-day regular exercise training could increase the number of EPC colonies and decrease homocysteine levels. Moreover, an inverse correlation was observed between small-endothelial progenitor cell colony-forming units (small-EPC-CFUs) and plasma homocysteine levels in healthy men (r=-0.8125, p=0.047). We found that regular exercise training could increase the number of EPC-CFUs and decrease homocysteine levels, thus decreasing the cardiovascular disease risk in men.

Tyrosine Kinase Inhibitors in Ph+ Chronic Myeloid Leukemia Therapy: a Review

  • Shah, Krupa;Parikh, Sonia;Rawal, Rakesh
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3025-3033
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    • 2016
  • Chronic myeloid leukaemia (CML) is a clonal myeloproliferative hematopoietic stem cell disorder. Deregulated BCR-ABL fusion tyrosine kinase activity is the main cause of CML disease pathogenesis, making BCR-ABL an ideal target for inhibition. Current tyrosine kinase inhibitors (TKIs) designed to inhibit BCR-ABL oncoprotein activity, have completely transformed the prognosis of CML. Interruption of TKI treatment leads to minimal residual disease reside (MRD), thought to reside in TKI-insensitive leukaemia stem cells which remain a potential reservoir for disease relapse. This highlights the need to develop new therapeutic strategies for CML either as small molecule master TKIs or phytopharmaceuticals derived from nature to achieve chronic molecular remission. This review outlines the past, present and future therapeutic approaches for CML including coverage of relevant mechanisms, whether ABL dependent or independent, and epigenetic factors responsible for developing resistance against TKIs. Appearance of mutant clones along the course of therapy either pre-existing or induced due to therapy is still a challenge for the clinician. A proposed in-vitro model of generating colony forming units from CML stem cells derived from diagnostic samples seems to be achievable in the era of high throughput technology which can take care of single cell genomic profiling.

Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells (Sca-1+골수조혈세포에서 JAK2/STAT5/GATA-1 신호전달 경로를 통한 다채, 도두 그리고 두 조합물에 의한 조혈증진 조절에 관한 연구)

  • Kim, Kunhoae;Kim, Seung-Hyung;Cho, In-Sik;Kim, Han-Young;Kim, Dong-Seon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.7-16
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    • 2013
  • Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

The build angle of 3D printing denture base resin on candida albicans adhesion. (의치상레진의 3D 프린팅 출력 각도가 Candida albicans의 부착에 미치는 영향)

  • Park, Su-Jung;Song, Young-Gyun
    • The Journal of the Korean dental association
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    • v.58 no.1
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    • pp.19-26
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    • 2019
  • Purpose: The purpose of this study is to compare the adhesion of Candida albicans according to build angle in 3D printing denture base resin. Methods: The 3D printing was performed by setting the build angle of the disk type specimen designed by CAD program at 0 degree, 30 degrees, 60 degrees, and 90 degrees. Surface roughness was measured using a non-contact 3D microsurface profiler. The specimens were incubated in Candida albicans suspension for 24 hours. The attached Candida albicans were detached by cell scraper. The suspension of detached C. albicans was serially diluted and plated on Trypticase soy broth. After 48 hours of incubation, total colony forming unit was counted. Results: There was no significant difference in surface roughness(Sa) between the test groups, but the interlayer boundary was observed. There was no statistically significant difference in total colony forming units of Candida albicans between the test groups. Conclusion: There was no difference in the average surface roughness and adhesion of Candida albicans between the specimens. It is considered that the setting of the build angle should be set considering the accuracy or strength rather than the roughness of the surface.

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A comparative study of three different viability tests for chemically or thermally inactivated Escherichia coli

  • Park, Seon Yeong;Kim, Chang Gyun
    • Environmental Engineering Research
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    • v.23 no.3
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    • pp.282-287
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    • 2018
  • Three different methods of bacterial viability monitoring were compared to detect chemically or thermally inactivated Escherichia coli. Direct colony enumeration, live/dead bacterial cell staining with a fluorescent dye, and the dehydrogenase activity assay were compared with respect to their ease of use and time required to perform the three different tests. The green (live cell)/red (dead cell) ratio obtained from the fluorescent bacterial cell staining approach showed a linear relationship with the colony forming units; the result obtained with dehydrogenase was similar to those. The sensitivity of the monitoring methods to detect bacterial deactivation varied with different disinfection conditions. After thermal treatment, the sensitivity of the staining approach was lower, while that of the dehydrogenase activity assay was the highest. After chemical treatment, the sensitivity of detection for both methods was similar.

Hydrogel Ocular Inserts for the Treatment of Infectious Bovine Keratoconjunctivitis

  • Ryoo, Je-Phil;Greer, R.T.;Rosenbusch, R.F.
    • Journal of Pharmaceutical Investigation
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    • v.22 no.3
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    • pp.35-47
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    • 1992
  • Hydrogel coated ring shaped ocular inserts (containing the antibiotic, tylosin tartrate) were used in an evaluation of the effectiveness of polymeric ocular drug release devices for treating infectious bovine keratoconjunctivitis. The in vivo experiments represent the first experiments using hydrogel ocular inserts containing an antibiotic for treating infectious bovine keratoconjunctivitis. In the infection tests, ten calves. were challenged with $2.4{\times}10^8{\sim}1.6{\times}10^9$ Moraxella bovis (a bacterium) colonies per eye following two ten minute ultraviolet radiation eye preconditioning exposures. Ninety five percent of the eyes (19 of 20 eyes) were successfully infected by this method. All infected eyes were monitored for the presence of the bacteria quantitiatively, and clinical observations were made for 14 days. The test was performed by three consecutive steps: 1) inoculation with 2 ultraviolet (UV) radiations, 2) growth of bacterial colonies and 3) treatment with medicated ring-shaped devices. The first. bacteriological measurements after 2 UV exposures were performed at day 3 of the tests. At day 7 after inoculation of both eyes of a calf with M. bovis, a medicated or a non-medicated ring-shaped device was inserted into each eye of a calf. The eye receiving the non-medicated ring was taken as a control for comparison with the eye that received a medicated ring. During the next 7 day period following a medicated ring insertion, the number of bacteria in the treated eyes dropped dramatically to negligible levels (0 to 30 colony forming units/swab), while the control eyes which received a non-medicated ring still exhibited a relatively high number of bacteria ($10^3\;to\;10^6$ colony forming units/swab). The number of bacteria was significantly reduced by the antibiotic released from the medicated ocular insert.

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