• 제목/요약/키워드: Single cell injection

검색결과 132건 처리시간 0.04초

A Disposable BioChip for Single Cell Manipulation

  • Yoon, Euisik
    • 한국반도체및디스플레이장비학회:학술대회논문집
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    • 한국반도체및디스플레이장비학회 2004년도 International Symposium
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    • pp.1-15
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    • 2004
  • o Various microfluidic components including mixromixers and micropumps have been developed for disposable biochip applications. o Single cell capturing, positioning and nanoliter drug injection chip has been demostrated. o Multi-channel, two-dimensional micro-well array has been fabricated and cell capturing and specific reagent injection have been performed.

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Increasing injection frequency enhances the survival of injected bone marrow derived mesenchymal stem cells in a critical limb ischemia animal model

  • Kang, Woong Chol;Oh, Pyung Chun;Lee, Kyounghoon;Ahn, Taehoon;Byun, Kyunghee
    • The Korean Journal of Physiology and Pharmacology
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    • 제20권6호
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    • pp.657-667
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    • 2016
  • Critical limb ischemia (CLI) is one of the most severe forms of peripheral artery diseases, but current treatment strategies do not guarantee complete recovery of vascular blood flow or reduce the risk of mortality. Recently, human bone marrow derived mesenchymal stem cells (MSCs) have been reported to have a paracrine influence on angiogenesis in several ischemic diseases. However, little evidence is available regarding optimal cell doses and injection frequencies. Thus, the authors undertook this study to investigate the effects of cell dose and injection frequency on cell survival and paracrine effects. MSCs were injected at $10^6$ or $10^5$ per injection (high and low doses) either once (single injection) or once in two consecutive weeks (double injection) into ischemic legs. Mice were sacrificed 4 weeks after first injection. Angiogenic effects were confirmed in vitro and in vivo, and M2 macrophage infiltration into ischemic tissues and rates of limb salvage were documented. MSCs were found to induce angiogenesis through a paracrine effect in vitro, and were found to survive in ischemic muscle for up to 4 weeks dependent on cell dose and injection frequency. In addition, double high dose and low dose of MSC injections increased vessel formation, and decreased fibrosis volumes and apoptotic cell numbers, whereas a single high dose did not. Our results showed MSCs protect against ischemic injury in a paracrine manner, and suggest that increasing injection frequency is more important than MSC dosage for the treatment CLI.

백서 복직근피판의 허혈-재관류 손상에 대한 히스타민 차단제의 효과 (The Effects of Anti-Histamine and Mast Cell Stabilizer against Ischemia-Reperfusion Injury to TRAM Flap in Rat)

  • 강상윤;정윤규;심정연
    • Archives of Plastic Surgery
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    • 제33권6호
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    • pp.742-747
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    • 2006
  • Purpose: The purpose of this study was to evaluate the role of mast cell and histamine as typical product of mast cell in ischemia-reperfusion injury of muscle flap using H2 receptor blocker and mast cell stabilizer. Methods: Thirty-five Sprague-Dawley rats weighing 250-300 gm were divided into four groups; Group I: Control group without ischemia, Group II: Normal saline injection group with ischemia, Group III: Cimetidine injection group with ischemia, Group IV: Sodium cromoglycate injection group with ischemia. Well established single pedicled transverse rectus abdominis musculocutaneous(TRAM) flap was designed in all rats and were rendered ischemia by clamping the artery for 150 minutes. All injections were applied intramuscular around gluteal area 30 minutes before reperfusion. The flap survival was evaluated at 7 days after operation. Neutrophil counts and mast cell counts were evaluated 24 hours after reperfusion. Results: The difference of skin flap survival between control group and cimetidine injection group was not significant. In the normal saline injection group flap survival was markedly decreased compared to that of control group. The muscle flap survival was similar to the results of skin flap survival. The neutrophil counts were significantly decreased in control group and sodium cromoglycate injection group than normal saline injection group. The mast cell counts were significantly decreased in cimetidine injection group and control group than both normal saline injection and sodium cromoglycate injection groups. The protective effect of sodium cromoglycate was not seen in the skin flap, but the muscle flaps showed protective effects of sodium cromoglycate compared to normal saline injection group. Conclusions: It is suggests that commonly used antihistamine(H2 receptor blocker) has protective effect against ischemia-reperfusion injury to skin and muscle flaps by reducing neutrophil and mast cell. The mast cell stabilizer was not effective for skin flap but, possibly, for muscle flap.

Effects on hematology and blood biochemistry profile of intramuscular meloxicam injection in Brahminy kite and Barn owl

  • Ratiwan Sitdhibutr;Raveewan Ploypan;Sirawit Subaneg;Chaiyan Kasorndorkbua
    • Journal of Veterinary Science
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    • 제24권3호
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    • pp.43.1-43.8
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    • 2023
  • Background: Meloxicam is used widely for exotic animal analgesia, but its toxicity in common raptor species in Thailand is unclear. Objectives: This study evaluated the single-dose effect of intramuscular meloxicam in common raptor species in Thailand for short-term and long-term periods. Methods: Twenty-two raptors were administered a single 1 mg/kg dose of meloxicam individually via intramuscular injection. The following were evaluated: clinical appearance, body weight, body condition score, body temperature, fecal appearance, complete blood cell count, and biochemistry panel before (day 0) and after the injection (1, 7, and 30 days). The collected samples were categorized into three groups: Brahminy kite (Haliastur indus) (n = 10), adult Barn owl (Tyto javanica) (n = 4), and juvenile Barn owl (n = 8). Results: None of the raptors in the study groups showed any abnormalities. The hematological profiles were significantly different in the short-term period (day 1 and day 7). The creatinine, aspartate aminotransferase, and creatinine kinase increased in several groups. On the other hand, the packed cell volume decreased in the Brahminy kite and juvenile Barn owl groups. According to the findings, an intramuscular injection of 1 mg/kg meloxicam affected the blood biochemistry panel of the muscle, but the affected raptors recovered within one week. Conclusions: An intramuscular injection of meloxicam at a single 1 mg/kg dose in Brahminy kites and Barn owls was not associated with the morbidity, hepatotoxicity, gastrointestinal toxicity, and nephrotoxicity in the short- and long-term periods.

In Vivo Spinal Distribution of Cy5.5 Fluorescent Dye after Injection via the Lateral Ventricle and Cisterna Magna in Rat Model

  • Lee, Kee-Hang;Nam, Hyun;Won, Jeong-Seob;Hwang, Ji-Yoon;Jang, Hye Won;Lee, Sun-Ho;Joo, Kyeung Min
    • Journal of Korean Neurosurgical Society
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    • 제61권4호
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    • pp.434-440
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    • 2018
  • Objective : The purpose of this study was to find an optimal delivery route for clinical trials of intrathecal cell therapy for spinal cord injury in preclinical stage. Methods : We compared in vivo distribution of Cy5.5 fluorescent dye in the spinal cord region at various time points utilizing in vivo optical imaging techniques, which was injected into the lateral ventricle (LV) or cisterna magna (CM) of rats. Results : Although CM locates nearer to the spinal cord than the LV, significantly higher signal of Cy5.5 was detected in the thoracic and lumbar spinal cord region at all time points tested when Cy5.5 was injected into the LV. In the LV injection Cy5.5 signal in the thoracic and lumbar spinal cord was observed within 12 hours after injection, which was maintained until 72 hours after injection. In contrast, Cy5.5 signal was concentrated at the injection site in the CM injection at all time points. Conclusion : These data suggested that the LV might be suitable for preclinical injection route of therapeutics targeting the spinal cord to test their treatment efficacy and biosafety for spinal cord diseases in small animal models.

Development of PCR based approach to detect potential mosaicism in porcine embryos

  • Cho, Jongki;Uh, Kyungjun;Ryu, Junghyun;Fang, Xun;Bang, Seonggyu;Lee, Kiho
    • 한국동물생명공학회지
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    • 제35권4호
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    • pp.323-328
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    • 2020
  • Direct injection of genome editing tools such as CRISPR/Cas9 system into developing embryos has been widely used to generate genetically engineered pigs. The approach allows us to produce pigs carrying targeted modifications at high efficiency without having to apply somatic cell nuclear transfer. However, the targeted modifications during embryogenesis often result in mosaicism, which causes issues in phenotyping founder animals and establishing a group of pigs carrying intended modifications. This study was aimed to establish a genomic PCR and sequencing system of a single blastomere in the four-cell embryos to detect potential mosaicism. We performed genomic PCR in four individual blastomeres from four-cell embryos. We successfully amplified target genomic region from single blastomeres of 4-cell stage embryo by PCR. Sanger sequencing of the PCR amplicons obtained from the blastomeres suggested that PCR-based genotyping of single blastomere was a feasible method to determine mutation type generated by genome editing technology such as CRISPR/Cas9 in early stage embryos. In conclusion, we successfully genotyped single blastomeres in a single 4-cell stage embryo to detect potential mosaicism in porcine embryos. Our approach offers a simple platform that can be used to screen the prevalence of mosaicism from designed CRISPR/Cas9 systems.

SONOS 구조를 갖는 멀티 비트 소자의 프로그래밍 특성 (Programming Characteristics of the multi-bit devices based on SONOS structure)

  • 안호명;김주연;서광열
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2003년도 하계학술대회 논문집 Vol.4 No.1
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    • pp.80-83
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    • 2003
  • In this paper, the programming characteristics of the multi-bit devices based on SONOS structure are investigated. Our devices have been fabricated by $0.35\;{\mu}m$ complementary metal-oxide-semiconductor (CMOS) process with LOCOS isolation. In order to achieve the two-bits per cell operation, charges must be locally trapped in the nitride layer above the channel near the junction. Channel hot electron (CHE) injection for programming can operate in multi-bit using localized trap in nitride film. CHE injection in our devices is achieved with the single power supply of 5 V. To demonstrate CHE injection, substrate current (Isub) and one-shot programming curve were investigated. The multi-bit operation which stores two-bit per cell is investigated with a reverse read scheme. Also, hot hole injection for fast erasing is used. Due to the ultra-thin gate dielectrics, our results show many advantages which are simpler process, better scalability and lower programming voltage compared to any other two-bit storage flash memory. This fabricated structure and programming characteristics are shown to be the most promising for the multi-bit flash memory.

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Real-Time Force Sensing in the Envelope of Zebrafish Egg during Micropipette Penetration

  • Yun, Seok;Kim, Deok-Ho;Kim, Byung-Kyu;Lee, Sang-Ho;Park, Gwi-Tae
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2003년도 ICCAS
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    • pp.2451-2456
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    • 2003
  • In biological cell manipulation, manual thrust or penetration of an injection pipette into an egg is currently performed by a skilled operator, relying only on visual feedback information. Massive load of various micro injection of either genes, fluid or cells in the postgenomic era calls a more reliable and automatic micro injection system that can test hundreds of genes or cell types at a single experiment. We initiated to study cellular force sensing in zebrafish eggs as the first step for the development of a more controllable micro injection system by any inexperienced operator. Zebrafish eggs at different developmental stages were collected and an integrated biomanipulation system was employed to measure cellular force during penetrating the egg envelope, the chorion. First of all, the biomanipulation system integrated with cellular force sensing instrument is implemented to measure the penetration force of cell membranes and characterize mechanical properties of zebrafish embryo cells. Furthermore, implementation of cellular force sensing system and calibration are presented. Finally, the cellular force sensing of penetrating cell membranes at each developmental stages was experimentally performed. The results demonstrated that the biomanipulation system with force sensing capability can measure cellular force at real-time while the injection operation is undergoing. The magnitude of the measured force was in the range of several hundreds of uN. The precise real-time measurement should provide the first step forwards for the development of an automatic and reliable injection system of various materials into biological cells.

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