• Title/Summary/Keyword: Isolation rate

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Presence of Leukemia-maintaining Cells in Differentiation-resistant Fraction of K562 Chronic Myelogenous Leukemia (만성 골수성 백혈병 K562세포의 분화 내성 분획에서 백혈병 유지 세포의 동정)

  • Lee, Hong-Rae;Kim, Mi-Ju;Ha, Gahee;Kim, So-Jung;Kim, Sun-Hee;Kang, Chi-Dug
    • Journal of Life Science
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    • v.23 no.2
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    • pp.197-206
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    • 2013
  • The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.

Evaluation of Control Pollination Efficiency and Management Status in Control Pollinated Progeny Populations of Pinus densiflora using Pedigree Analysis based on Microsatellite Markers (소나무 인공교배 차대집단에서 Microsatellite marker 혈통분석을 이용한 인공교배 효율 및 관리상태 평가)

  • Tae-Lim Yeo;Jihun Kim;Dayoung Lee;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.2
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    • pp.157-172
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    • 2023
  • Controlled pollination (CP) is an important method in tree breeding programs because CP quickly generates desirable genotypes and rapidly maximizes genetic gains. However, few studies have evaluated the efficiency and success rate of CP in the breeding program of Pinus densiflora. To evaluate CP and the management of control pollinated progenies, we used 159 individuals in CB2 × KW40 or KW40 × CB2 populations that were established in 2015. After genotyping microsatellite loci, we estimated whether the number of primers was sufficient or not. Then, we performed pedigree analysis. The result showed that the number of primers was sufficient. By pedigree analysis, we found out that 60 of 159 individuals had been generated by the mating between CB2 and KW40. In the maternity analysis, there was evidence to indicate the possibility of management problems. Therefore, we excluded 54 individuals and repeated the pedigree analysis. In the second analysis, 47 of 105 individuals were generated by the mating between CB2 and KW40. To increase the efficiency of CP in tree breeding programs, several precautions are required. It is necessary to identify the exact clone names of the mother and father trees. In addition, CP processes should be performed properly, including deciding on the schedule of CP and the isolation of female strobili or flowers. Finally, the monitoring of hybrid progenies management after mating is important. Molecular markers should be used to identify the clone names of the mother and father trees and for monitoring post hoc management. This study provides a reference for tree breeding programs for the future control pollination of pine species.

Isolation and Characterization of the Indigenous Microalgae Chlamydomonas reinhardtii K01 as a Potential Resource for Lipid Production and Genetic Modification (지질생산 및 유전자 조작의 잠재적 자원으로서의 토착 미세조류 Chlamydomonas reinhardtii K01의 분리 및 특성)

  • Kim, Eun-Kyung;Cho, Dae Hyun;Suh, Sang-Ik;Lee, Chang-Jun;Kim, Hee-Sik;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.32 no.3
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    • pp.202-209
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    • 2022
  • The green alga Chlamydomonas reinhardtii, a unicellular haploid eukaryote, has long been used by researchers and industries as a cell factory to produce high value-added microalgae substances using genetic modification. Microalga K01, presumed to be Chlamydomonas, was isolated from 12 freshwater samples from the Chungcheong and Jeolla regions to replace C. reinhardtii, an introduced species currently used in most basic and industrial research. The isolated K01 strain was identified as C. reinhardtii through morphological and phylogenetic studies of the 18S rDNA gene sequence (NCBI accession number KC166137). The growth and lipid content of the isolated C. reinhardtii K01 were compared with three wild and four mutant strains in TAP medium, and it was found that the K01 strain could produce 1.74×107 cells/ml by the third day of culture. The growth rate of C. reinhardtii K01 was 1.5 times faster than UTEX2244, which showed the highest number of cells (1.20×107 cells/ml) among the compared strains. The lipid content of the isolated C. reinhardtii K01 (20.67%) was similar to those of the wild strains, although the fatty acid oleate C18:1 was not detected in the isolated strain but was identified in the seven others. The cell density of the isolated strain increased to 0.87 g/l during a six-day culture in BG11 medium, where nitrate (NaNO3) was introduced as a nitrogen source, while the seven acquired strains showed almost no cell proliferation.

Investigation of Microbial Contamination in the Raw Materials of Meal Kits (간편조리세트 원재료의 미생물 오염도 조사)

  • Hyun-Kyung Lee;Young-Sook Do;Min-Jung Park;Kyoung Suk Lim;Seo-In Oh;Jeong-Hwa Lim;Hyun-Soo Kim;Hyun-Kyung Ham;Yeo-Jung Kim;Myung-Jin Lee;Yong-Bae Park
    • Journal of Food Hygiene and Safety
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    • v.39 no.2
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    • pp.109-117
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    • 2024
  • This study investigated the microbial contamination of agricultural, livestock, and marine ingredients in 55 meal kits distributed across Gyeonggi-do, South Korea. Of the 55 meal kits, 48 contained agricultural ingredients, 43 contained livestock ingredients, and 16 contained marine ingredients. The detection rate of the total aerobic bacteria in the agricultural, livestock, and marine products was 100%. The average numbers of the total aerobic bacteria were 6.57 log colony-forming units (CFU)/g in the agricultural products, 4.60 log CFU/g in the livestock products, and 5.47 log CFU/g in the marine products. The coliform detection rates in the agricultural, livestock, and marine products were 81.25%, 69.77%, and 43.75%, respectively. The average numbers of coliforms were 2.83 log CFU/g in the agricultural products, 1.34 log CFU/g in the livestock products, and 1.12 log CFU/g in the marine products. Escherichia coli was detected in 13 livestock products (30.23%), with levels ranging from 0.70 to 2.36 log CFU/g. Contrastingly, E. coli was detected in only one marine product (6.25%) and was not detected in any agricultural products. The detection rates of fungi in agricultural, livestock, and marine products were 97.92%, 93.02%, and 93.75%, respectively. The average numbers of fungi were 3.82 log CFU/g for the agricultural products, 2.92 log CFU/g for the livestock products, and 2.82 log CFU/g for the marine products. The isolation rates of foodborne pathogens from the agricultural, livestock, and marine products were 35.42%, 37.21%, and 31.25%, respectively. Forty-five foodborne pathogens of seven species, including Bacillus cereus and Salmonella spp., were isolated from the raw materials of the agricultural, livestock, and marine products in 55 meal kits. To prevent foodborne diseases caused by meal kits, it is necessary to focus on washing, heating, and preventing cross-contamination during cooking.