• 제목/요약/키워드: high cell density

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Insertional mutations exhibiting high cell-culture density HCD phenotypes are enriched through continuous subcultures in Chlamydomonas reinhardtii

  • Thung, Leena;He, Jing;Zhu, Qingling;Xu, Zhenyu;Liu, Jianhua;Chow, Yvonne
    • ALGAE
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    • v.33 no.1
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    • pp.127-141
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    • 2018
  • Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.

Differential synapse density between Purkinje cell dendritic spine and parallel fiber varicosity in the rat cerebellum among the phylogenic lobules

  • Hyun-Wook Kim;Seung Hak Oh;Se Jeong Lee;Ji eun Na;Im Joo Rhyu
    • Applied Microscopy
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    • v.50
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    • pp.6.1-6.6
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    • 2020
  • The cerebellum is a region of the brain that plays an important role in motor control. It is classified phylogenetically into archicerebellum, paleocerebellum and neocerebellum. The Purkinje cells are lined in a row called Purkinje cell layer and it has a unique dendritic branches with many spines. The previous study reported that there is a difference of synapse density according to the lobules based on large two-dimensional data. However, recent study with high voltage electron microscopy showed there was no differences in dendritic spine density of the Purkinje cell according to its phylogenetic lobule. We analyzed Purkinje cell density in the II, VI and X lobules by stereological modules and synaptic density was estimated by double disector based on Purkinje cell density in the molecular layer of each lobule. The results showed that there was significant difference in the Purkinje cell density and synapse number according to their phylogenetic lobules. The number of Purkinje cell in a given volume was larger in the archicerebellum, but synapse density was higher in the neocerebellum. These data suggest that cellular and synaptic organization of the Purkinje cell is different according to their phylogenetic background.

Design of flow path with 2 inlet and outlets to improve cell performance and prevent cell degradation in Solid Oxide Fuel Cell (SOFC 셀 성능 향상 및 수명 저하 방지를 위한 입구와 출구 2개의 유로 설계)

  • Kim, Dongwoo;Yeom, Eunseop
    • Journal of the Korean Society of Visualization
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    • v.19 no.2
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    • pp.56-62
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    • 2021
  • Solid oxide fuel cells (SOFCs) is the high efficiency fuel cell operating at high temperatures ranging from 700-1000℃. Design of the flow paths of the fuel and air in SOFCs is important to improve cell performance and prevent cell degradation. However, the uneven distribution of current density in the traditional type having one inlet and outlet causes cell degradation. In this regard, the parallel flow path with two inlet and outlets was designed and compared to the traditional type based on computational fluid dynamics (CFD) simulation. To check the cell performance, hydrogen distribution, velocity distribution and current density distribution were monitored. The results validated that the parallel designs with two inlets and outlets have a higher cell performance compared to the traditional design with one inlet and outlet due to a larger reaction area. In case of uniform-type paths, more uniform current density distribution was observed with less cross-sectional variation in flow paths. In case of contracted and expanded inflow paths, significant improvement of performance and uniform current density was not observed compared to uniform parallel path. Considering SOFC cell with uniform current density can prevent cell degradation, more suitable design of SOFC cell with less cross-sectional variation in the flow path should be developed. This work can be helpful to understand the role of flow distribution in the SOFC performance.

High-Cell-Density Fed-Batch Culture of Saccharomyces cerevisiae KV-25 Using Molasses and Corn Steep Liquor

  • Vu, Van Hanh;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1603-1611
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    • 2009
  • High-cell-density cultivation of yeast was investigated using the agricultural waste products corn steep liquor (CSL) and molasses. The Saccharomyces cerevisiae KV-25 cell mass was significantly dependent on the ratio between C and N sources. The concentrations of molasses and CSL in the culture medium were statistically optimized at 10.25% (v/v) and 16.87% (v/v), respectively, by response surface methodology (RSM). Batch culture in a 5-l stirred tank reactor using the optimized medium resulted in a cell mass production of 36.5 g/l. In the fed-batch culture, the feed phase was preceded by a batch phase using the optimized medium, and a very high dried-cell-mass yield of 187.63 g/l was successfully attained by feeding a mixture of 20% (v/v) molasses and 80% (v/v) CSL at a rate of 22 ml/h. In this system, the production of cell mass depended mainly on the agitation speed, the composition of the feed medium, and the glucose level in the medium, but only slightly on the aeration rate.

High Cell Density Cultivation of Pseudomonas putida BM01 Using Glucose

  • Kim, Guk Jin;In Young Lee;Dae Keon Choi;Sung Chul Yoon;Young Hoon Park
    • Journal of Microbiology and Biotechnology
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    • v.6 no.3
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    • pp.221-224
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    • 1996
  • Pseudomonas putida BM01 was grown efficiently on glucose as the sole carbon source with a supply of a nitrogen source in pH-stat mode using a low setpoint limit. A final cell concentration of 100 g/l was obtained in 30 h of fed-batch cultivation by controlling glucose concentration within the range of 5-20 g/l and maintaining dissolved oxygen tension above 10$%$ saturation using pure oxygen. This high cell density culture technique is believed highly useful for the production of poly(3-hydroxyalkanoates) by this strain.

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High Cell Density Culture of Bifidobacterium longum by Cross-flow Filtration (Cross-flow filtration에 의한 Bifidobacterium longum의 고농도 배양)

  • Lee, Myong-Suk;Park, Yun-Hee
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.18-22
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    • 1997
  • The conditions for production of high cell density of Bifidobacterium longum were investigated and the cross-flow filtration system was used to remove the inhibitory metabolites, lactic acid and acetic acid. The maximum cell growth was observed with glucose as carbon source at the concentration of 50 g/l at $37^{\circ}C$ with the initial pH 6.5. When B. longum was cultured in a cross-flow filtration system, the maximum cell growth was observed at a dilution rate(D) of $0.31\;h^{-1}$ and the dry cell weight was 16.4 g/l($3.5{\times}10^{10}\;cell/ml$), which was about four times higher than that obtained in the batch culture with pH control.

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Lipid and Lipase Distribution on Endosperm Cell of Panax ginseng Seed for the Electron Microscope (전자현미경을 이용한 인삼종자 배유세포내의 지질 및 지질가수분해 효소의 분포)

  • 유성철;노미전
    • Journal of Ginseng Research
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    • v.16 no.2
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    • pp.129-137
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    • 1992
  • This study was carried out to investigate the localization of lipids and lipase activity with lipid staining and cytochemical technique in endosperm cells of Panax ginseng C.A. Meyer seed. In endosperm cells of indehiscent seed, protein bodies facing the umbiliform layer are different in electron density during the various degraded processes. Gradually, protein matrix near the cell wall was lysed and electron lucent inclusions appeared on umbiliform layer. The protein body with high electron density and the spherosome with low electron density were observed in endosperm cells. As a result of lipid staining, electron density of spherosome is more intense than those of the protein matrix within the protein body in endosperm cells of indehiscent seed. Free spherical spherosomes within the umbiliform layer have a high electron density. The spherical spherosomes were more electron densed and were uniform in comparison with the cytoplasmic proteinaceous granules in endosperm cells of seed with red seed coat. The major component of spherosome was determined to be lipid. Lipase activity occurs in the spherosome and near the endosperm cell wall facing the umbiliform layer. Cytochemical reaction products of lipase were observed in the spherosome membrane and in the inner regions of spherosome. After protein bodies were digested, lipase activities were observed in free spherosomes and near the cell wall of endosperm cells. Umbiliform layer composing of fibrillized wall and digested materials of the endosperm cell showed a little lipase reaction products.

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A High-Density 64k-Bit One-Time Programmable ROM Array with 3-Transistor Cell Standard CMOS Gate-Oxide Antifuse

  • Cha, Hyouk-Kyu;Kim, Jin-Bong;Lee, Kwy-Ro
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.4 no.2
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    • pp.106-109
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    • 2004
  • A high-density 3-transistor cell one-time programmable (OTP) ROM array using standard CMOS Gate-Oxide antifuse (AF) is proposed, fabricated, and characterized with $0.18{\mu}m$ CMOS process. The proposed non-volatile high-density OTP ROM is composed of an array of 3-T OTP cells with the 3-T consisting of an nMOS AF, a high voltage (HV) blocking transistor, and a cell access transistor, all compatible with standard CMOS technology.

Cell separation from high density culture broths of Alcaligenes eutrophus by using Al-based coagulants (Alcaligens eutrohus 고농도 배양액으로부터 알루미늄(Al)계 응집제를 이용한 세포분리)

  • 조경숙;류희욱;정현우;곽종운;장용근
    • KSBB Journal
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    • v.13 no.3
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    • pp.272-278
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    • 1998
  • Cell recovery from high cell density broths of Alcaligenes eutrophus by pretreatment with aluminum-based coagulants such as aluminum sulfate, polyaluminum hydrooxide chloride silicate (PACS), and polyaluminum hydrooxide chloride (Hi-PAX) was carried out. Cells coagulated with coagulants could be successfully recovered above 95-99% by centrifugation or filtration. The optimum initial pH of fermentation broths for cell recovery was in the range of 10 to 12. Optimum coagulants dosage for cell recovery increased with increasing of cell concentrations (21-160 g/L). The optimum coagulant dosages to recover cells with more than 95% cell recovery by centrifugation for the cell concentrations ranged 21-160 g/L were as follows: aluminum sulfate, 416-1708 mg Al/L; PACS, 211-826 mg Al/L; Hi-PAX, 320-960 mg Al/L. At optimum conditions for the coagulation of cells, centrifugal forces for 95% of cell recovery were dependent on the cell concentration. The centrifugal forces at 82 g/L and 160 g/L of cell concentration were only 45${\times}$g and 1600${\times}$g, respectively.

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High Cell Density Cultivation of Pseudomonas oleovorans for the Production of Poly(3-Hydroxyalkanoates)

  • Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.51-53
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    • 1996
  • Fed-batch culture of Pseudomonas oleovorans was carried out for the production of medium-chain-length polyhydroxyalkanoates (MCL-PHAs) using octanoate as a carbon source. Octanoate and the salt solution containing ammounium sulfate and magnesium sulfate were intermittently fed in the course of fermentation. Cell mass and PHA concentrations of 42.8 and 16.8g/L, respectively, could be obtained in 40 h. The PHA content and the PHA productivity were 39.2% and 0.42 g PHA/L-h, respectively. The yields of cell mass and PHA were 0.71 g dry cell mass/g octanoate and 0.28g PHA/g octanoate, respectively. Therefore, octanoate can be used for the production of MCL-PHAs to a high concentration with high productivity.

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