• Title/Summary/Keyword: Bank of Human Cell

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STANDARD OPERATING PROCEDURES OF HARD TISSUES SUCH AS FEMORAL HEAD, ALLOGRAFTS OBTAINED FROM LIVING DONORS (생존 기증자로부터 채취된 경조직(대퇴골두 등)의 조직은행 술식)

  • Lee, Eun-Young;Kim, Kyung-Won;Um, In-Woong;Ryu, Ju-Youn
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.5
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    • pp.406-413
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    • 2004
  • Progress in medical science and cell biology has resulted in the transplantation of human cells and tissues from on human into another, facilitating reproduction and the restoration of form and function, as well as enhancing the quality of life. For more than 40 years, society has recognized the medical and humanitarian value of donation and transplanting organs and tissues. The standard operating procedures of hard tissues reflect the collective expertise and conscientious efforts of tissue bank professionals to provide a foundation for the guidance of tissue banking activities. Procurement of allograft tissues from surgical bone donors is a part of tissue banking. During the past decades the use of bone allografts has become widely accepted for the filling of skelectal defects in a variety of surgical procedures. In particular in the field of orthopaedic and oral and maxillofacial surgery the demand for allografts obtained from either living or post-mortem donors has increased. Hospital-based tissue banks mainly retrieve allografts from living donors undergoing primary total hip replacement for osteoarthritis or hemi arthroplasty for hip fractures and orthgnatic surgery such as angle reduction. Although bone banks have existed for many years, the elements of organized and maintaining a hospital bone bank have not been well documented. The experience with a tissue bank at Korea Tissue Bank(KTB) between 2001 and 2004 provides a model of procurement, storage, processing, sterilization and documentation associated with such a facility. The following report describes the standard operating procedures of hard tissues such as femoral head obtained from living donors.

Distributions of HLA Microsatellite Markers and the Linkage Disequilibria between HLA and Microsatellites in Koreans (한국인에서 HLA 유전자 부위 내 Microsatellite 표지자의 분포와 HLA 대립유전자의 유전적 연관성)

  • Jang, Jung-Pil;Choi, Eun-Jeong;Yoon, Ho-Yeul;Choi, Hee-Baeg;Kim, Hee-Je;Cho, Byung-Sik;Min, Woo-Sung;Lee, Jong-Wook;Kim, Chun-Choo;Kim, Tai-Gyu
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.149-157
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    • 2007
  • Background: The microsatellites within human leukocyte antigen (HLA) region show considerable polymorphism and strong linkage disequilibrium (LD) with HLA alleles. These microsatellites have been used for genetic analysis including disease mapping to understand susceptibility to autoimmune and infectious diseases. Also, use of microsatellites has recently been proposed as an approach for identifying non-HLA markers within the HLA region that could function as transplantation determinants and for the selection of potential donors for transplantation. Methods: To analyse the frequency of five microsatellites in the Korean population, genotyping for polymorphisms at five microsatellites markers (BAT2, MIB, DQCAR, D6S105 and TNFd) within HLA region was performed on 143 healthy Korean controls. Results: The most frequent genotype shown in healthy Korean controls were BAT2 8 (153 bp, 42.7%), MIB 1 (326 bp, 40.6%), DQCAR 3 (188 bp, 38.5%), D6S105 7 (126 bp, 58.0%) and TNFd 3 (128 bp, 58.0%). And common two-loci haplotypes were found as MIB 1-HLA-B*62 (HF: 10.6%), MIB 6-HLA-B*44 (HF: 7.8%), DQCAR 3-HLA-DRB1*13 (HF: 8.5%), TNFd 5-HLA-B*62 (HF: 7.8%) and D6S105 7-HLA-A*02 (HF: 16.2%). Conclusion: These data might provide useful information on the microsatellites markers with HLA region in Korean population and be helpful in further defining the clinical impact of these microsatellites.

Effects of Musculoskeletal Transplant Foundation on Bone Formation in Human Fetal Osteoblasts (사람태아골모세포에 대한 근골격이식재의 골형성 유도에 관한 효과)

  • Park, Jae-young;Pi, Sung-Hee;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.449-459
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    • 2006
  • DFDBA(Decalcified freeze-dried bone allograft) is one of the allograft materials for periodontal bone regeneration. DFDBA provides an osteoconductive surface and osteoinductive factors. Therefore, DFDBA have been used successfully to regenerate the attachment apparatus during periodontal treatment. But recent studies was reported that wide variations in commercial bone bank preparations of DFDBA do exist, including the ability to induce new bone formation. DFDBA was experimental materials that was recovered, processed, tested, shipped and invoiced through Musculoskeletal Transplant Foundation. MTF(Musculoskeletal Transplant Foundation) is the world largest, non-profit, AATB(American Association of Tissue Banks) accredited tissue bank. The objective of this study was to determine the effects of serial dilutions of a DFDBA on human fetal osteoblastic cell proliferation and their potential to form and mineralize bone nodules. Human fetal osteoblastic cell line(hFOB 1.19) was cultured with DMEM and SSE($1{\mu}g/m{\ell}$,$10{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $1mg/m{\ell}$) at $34^{\circ}C$ with 5% CO2 in 100% humidity. Cell proliferation was significantly increased at $1mg/m{\ell}$, $100{\mu}g$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of DFDBA after 5 days incubation (p<0.05). Alkaline Phosphatase(ALP) level was significantly increased in $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of DFDABA(p<0.05). A quantified calcium accumulation was significantly increased at $1ng/m{\ell}$, $10ng/m{\ell}$ of MTF(p<0.05). These results indicated that DFDBA has an inductive effect on bone formation in vitro.

Comprehensive Analysis of Epstein-Barr Virus LMP2A-Specific CD8+ and CD4+ T Cell Responses Restricted to Each HLA Class I and II Allotype Within an Individual

  • Hyeong-A Jo;Seung-Joo Hyun;You-Seok Hyun;Yong-Hun Lee;Sun-Mi Kim;In-Cheol Baek ;Hyun-Jung Sohn;Tai-Gyu Kim
    • IMMUNE NETWORK
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    • v.23 no.2
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    • pp.17.1-17.16
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    • 2023
  • Latent membrane protein 2A (LMP2A), a latent Ag commonly expressed in Epstein-Barr virus (EBV)-infected host cells, is a target for adoptive T cell therapy in EBV-associated malignancies. To define whether individual human leukocyte antigen (HLA) allotypes are used preferentially in EBV-specific T lymphocyte responses, LMP2A-specific CD8+ and CD4+ T cell responses in 50 healthy donors were analyzed by ELISPOT assay using artificial Ag-presenting cells expressing a single allotype. CD8+ T cell responses were significantly higher than CD4+ T cell responses. CD8+ T cell responses were ranked from highest to lowest in the order HLA-A, HLA-B, and HLA-C loci, and CD4+ T cell responses were ranked in the order HLA-DR, HLA-DP, and HLA-DQ loci. Among the 32 HLA class I and 56 HLA class II allotypes, 6 HLA-A, 7 HLA-B, 5 HLA-C, 10 HLA-DR, 2 HLA-DQ, and 2 HLA-DP allotypes showed T cell responses higher than 50 spot-forming cells (SFCs)/5×105 CD8+ or CD4+ T cells. Twenty-nine donors (58%) showed a high T cell response to at least one allotype of HLA class I or class II, and 4 donors (8%) had a high response to both HLA class I and class II allotypes. Interestingly, we observed an inverse correlation between the proportion of LMP2A-specific T cell responses and the frequency of HLA class I and II allotypes. These data demonstrate the allele dominance of LMP2A-specific T cell responses among HLA allotypes and their intra-individual dominance in response to only a few allotypes in an individual, which may provide useful information for genetic, pathogenic, and immunotherapeutic approaches to EBV-associated diseases.

AN EXPERIMENTAL STUDY OF UNDEMINERALIZED FREEZE-DRIED HUMAN BONE (냉동건조 은행골의 개발 및 이의 실험적 연구)

  • Choung, Pil-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.18 no.1
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    • pp.164-174
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    • 1996
  • In order to develop the allogeneic bone implants instead of autogenous bone grafts for maxillofacial reconstruction, undemineralized freeze-dried human bone was processed. The freeze-dried human bone was implanted into the cranial and mandibular defects of the rabbits. The implants were evaluated clinically, roentgenographically and histomophometrically. And immunohistochemical evaluation of the implants was performed on the rat. The results were as follows : 1. When compared with control defects of $0.8{\times}0.8\;cm$, the implants on the rabbit defects displayed complete osseous bridging clinically and roentgenographically. Histomophometrically a minimal inflammatory cell infiltrate was present but the defects healed well clinically. 2. When compared with control grafts, the freeze-dried implants on the rat muscle displayed decreased antigenicity by immunohistochemical evaluation, due to freeze-drying process. 3. Undemineralized freeze-dried human bone in this study can be preserved as a bank bone in this study and seems to be applicable for clinical allogeneic bone grafts.

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HLA-restricted and Antigen-specific CD8+ T Cell Responses by K562 Cells Expressing HLA-A*0201

  • Yun, Sun-Ok;Sohn, Hyun-Jung;Yoon, Sung-Hee;Choi, Hee-Baeg;Kim, Tai-Gyu
    • IMMUNE NETWORK
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    • v.6 no.4
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    • pp.179-184
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    • 2006
  • Background: Identification of antigen-specific T cells has yielded valuable information on pathologic process and the disease state. Assays for quantification of inflammatory cytokines or lytic-granule molecules have been generally used to evaluate antigen specific T cell response, however their applicability have been hampered due to the limited source of autologous antigen-presenting target cells (APC). Methods: K562, a leukemic cell line deficient of human leukocyte antigen (HLA), was transfected with a gene encoding HLA-A*02 (K562/ A*02) and its function as stimulator cells in inducing activation of HLA-matched T cells was evaluated by IFN-${\gamma}$ enzyme linked immunospot (ELISPOT) assay. Results: The stable transfectant K562/ A*02 pulsed with HLA- A*02 restricted peptide could specifically induce IFN-${\gamma}$ secretion by CD8+ T cells compared to no detectable secretion by CD4+ T cells. However, CD56+ NK cells secreted IFN-${\gamma}$ in both K562/ A*02 with peptide and without peptide. The number of IFN-${\gamma}$ secreted CD8+ T cells was increased according to the ratio of T cells to K562 and peptide concentration. Formalin-fixed K562/ A*02 showed similar antigen presenting function to live K562/ A*02. Moreover, K562/ A*02 could present antigenicpeptide to not only A*0201 restricted CD8+ T cells but also CD8+ T cells from A*0206 donor. Conclusion: These results suggest that K562/ A*02 could be generally used as target having specificity and negligible background for measuring CD8+ T cell responses and selective use of K562 with responsder matched HLA molecules on its surface as APC may circumvent the limitation of providing HLA-matched autologous target cells.

Development of a Simple Method to Determine the Mouse Strain from Which Cultured Cell Lines Originated

  • Yoshino, Kaori;Saijo, Kaoru;Noro, Chikako;Nakamura, Yukio
    • Interdisciplinary Bio Central
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    • v.2 no.4
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    • pp.14.1-14.9
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    • 2010
  • Misidentification of cultured cell lines results in the generation of erroneous scientific data. Hence, it is very important to identify and eliminate cell lines with a different origin from that being claimed. Various methods, such as karyotyping and isozyme analysis, can be used to detect inter-species misidentification. However, these methods have proved of little value for identifying intra-species misidentification, and it will only be through the development and application of molecular biological approaches that this will become practical. Recently, the profiling of microsatellite variants has been validated as a means of detecting gene polymorphisms and has proved to be a simple and reliable method for identifying individual cell lines. Currently, the human cell lines provided by cell banks around the world are routinely authenticated by microsatellite polymorphism profiling. Unfortunately, this practice has not been widely adopted for mouse cells lines. Here we show that the profiling of microsatellite variants can be also applied to distinguish the commonly used mouse inbred strains and to determine the strain of origin of cultured cell lines. We found that approximately 4.2% of mouse cell lines have been misidentified; this is a similar rate of misidentification as detected in human cell lines. Although this approach cannot detect intra-strain misidentification, the profiling of microsatellite variants should be routinely carried out for all mouse cell lines to eliminate inter-strain misidentification.

Antiproliferative Properties of Methanolic Extract of Nigella sativa against the MDA-MB-231 Cancer Cell Line

  • Dilshad, Ahmad;Abulkhair, Omalkhair;Nemenqani, Dalal;Tamimi, Waleed
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5839-5842
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    • 2012
  • Breast cancer is the most commonly diagnosed cancer in women in the world and is one of the leading causes of death due to cancer. Health benefits have been linked to additive and synergistic combinations of phytochemicals in fruits and vegetables. Nigella sativa has been shown to possess anti-carcinogenic activity, inhibiting growth of several cancer cell lines in vitro. However, the molecular mechanisms of the anti-cancer properties of Nigella sativa phytochemical extracts have not been completely understood. Our data showed that Nigella sativa extracts significantly inhibited human breast cancer MDA-MB-231 cell proliferation at doses of $2.5-5{\mu}g/mL$ (P<0.05). Apoptotic induction in MDA-MB-231 cells was observed in a dose-dependent manner after exposure to Nigella sativa extracts for 48 h. Real time PCR and flow cytometry analyses suggested that Nigella sativa extracts possess the ability to suppress the proliferation of human breast cancer cells through induction of apoptosis.

Effects of Irradiated Frozen Allogenic Bone and Musculoskeletal Transplant Foundation on Bone Formation in Human Fetal Osteoblasts (사람 태아 골모 세포에 대한 냉동 동종골과 근골격이식재의 골형성 유도에 관한 효과)

  • Yoon, Ho-Sang;Pi, Sung-Hee;Yun, Hyeong-Geun
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.435-448
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    • 2006
  • The purpose of this study was to investigate the effects of ICB(Irradiated frozen allogenic bone, Rocky Mountain Tissue Bank, USA) and MTF(Decalcified freeze-dried bone allograft, Musculoskeletal Transplant Foundation, USA) on the cell proliferation and differentiation of human fetal osteoblasts. Human fetal osteoblasts (hFOB1) were cultured with $10\;ng/m{\ell}$of ICB and MTF. The negatvie control group was cultured with DMSO and positive control group was cultured with BMF ($2\;ng/m{\ell}$). MIT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examine the mineralization. Calcium accumulation was also evaluated. ICB and MTF did not increase the rate of the cellular proliferation of hFOB1s while they enhanced ALP and calcium accumulation. The expression of osteocalcin (OC) and bone silaloprotein (BSP) increased in hFOB1 treated with ICB and MTF ($10\;ng/m{\ell}$). These results suggest that ICB and MTF stimulate osteoblastic activity of the hFOBl.

Fruit Mediated Synthesis of Gold and Silver Nanoparticles Using Lycium chinense and Their Antimicrobial Activity

  • Chokkalingam, Mohan;Huo, Yue;Kang, Jong-Pyo;Mathiyalagan, Ramya;Kim, Yoen-Ju;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.94-94
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    • 2018
  • The gold (LC-AuNPs) and silver (LC-AgNPs) nanoparticles were rapidly synthesized by fruit extract of Lycium chinense within 1.15 and 25 min respectively in an eco-friendly way. The synthesized nanoparticles confirmed by relevant surface plasmon resonance peaks for gold and silver nanoparticles at 536 and 480 nm, respectively. FE-TEM results revealed that LC-AuNPs were 20-50 nm and LC-AgNPs were 50-100 nm. The maximum distribution of gold, silver elements and the crystallographic nature of synthesized were confirmed using EDX, elemental mapping and XRD. LC-AgNPs showed inhibitory activity against pathogenic microorganisms such as E. coli and S. aureus, whereas LC-AuNPs did not show inhibitory activity. The LC-AgNps nanoparticles exhibited significant cytotoxicity to human breast cancer MCF7 cell line and less cytotoxicity to non-diseased RAW264.7 (murine macrophage) cells whereas LC-AuNps showed minimal toxicity to both cell lines. In-depth research on this rapid, facile and greenery nanoparticles may play a potential role in biomedical applications.

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