• Title/Summary/Keyword: Lineage

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Analysis of Molecular Expression in Adipose Tissue-Derived Mesenchymal Stem Cells : Prospects for Use in the Treatment of Intervertebral Disc Degeneration

  • Jin, Eun-Sun;Min, Joongkee;Jeon, Sang Ryong;Choi, Kyoung Hyo;Jeong, Je Hoon
    • Journal of Korean Neurosurgical Society
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    • v.53 no.4
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    • pp.207-212
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    • 2013
  • Objective : Recent studies have shown encouraging progress toward the use of autogenic and allogenic mesenchymal stem cells (MSCs) to arrest, or even lead to partial regeneration in, intervertebral disc (IVD) degeneration. However, this technology is still in its infancy, and further development is required. The aim of this study was to analyze whether rat adipose-derived mesenchymal stem cells (ADMSC) can differentiate towards IVD-like cells after treatment with transforming growth factor ${\beta}3$ (TGF-${\beta}3$) in vitro. We also performed quantitative analysis of gene expression for ADMSC only, ADMSCs treated with TGF-${\beta}3$, and co-cultured ADMSCs treated with TGF-${\beta}3$. Methods : ADMSCs were sub-cultured to homogeneity and used in fluorocytometry assays for CD11, CD45, and CD90/Thy1. ADMSCs were differentiated in spheroid culture towards the chondrogenic lineage by the presence of TGF-${\beta}3$, dexamethasone, and ascorbate. We also co-cultured pure ADMSCs and nucleus pulposus cells in 24-well plates, and performed immunohistochemical staining, western blotting, and RT-PCR for quantitative analysis of gene expression. Results : Results of fluorocytometry were positive for CD90/Thy1 and negative for CD11 and CD45. TGF-${\beta}3$-mediated induction of ADMSCs led to the expression of the differentiation markers of intervertebral disc-like cells, such as aggrecan, collagen II, and sox-9. Co-cultured ADMSCs treated with TGF-${\beta}3$ showed higher expression of differentiation markers and greater extracellular matrix production compared with ADMSCs treated with TGF-${\beta}3$ alone. Conclusion : ADMSC treated with TGF-${\beta}3$ may be an attractive source for regeneration therapy in degenerative IVD. These findings may also help elucidate the pathologic mechanism of MSC therapy in the degeneration of IVD in vivo.

SNP-based Genetic Diversity and Relationships Analysis of the Korean Native Black Goat and Crossbred Goat (SNP 정보를 활용한 재래흑염소와 교잡종 염소의 유전적 다양성 및 유연관계 분석)

  • Lee, Sang-Hoon;Lee, Jinwook;Lee, Eun-Do;Kim, Seungchang;Lee, Sung-Soo;Kim, Kwan-Woo
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.11
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    • pp.102-108
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    • 2020
  • This study was conducted to investigate the genetic diversity and genetic taxonomic relationships between Korean native black goat (KNBG) populations and crossbred goats. The 45,658 common single nucleotide polymorphisms present in the KNBG strain and crossbred goat were used for the analysis. The expected and observed heterozygosity (which can be indicators of genetic diversity) were in the order of crossbred, Gyeongsang National University, Jangsu, then the Tongyeong strains. The variance component represents the degree of genetic diversity between groups. The highest variance (19.98 %) was between the Dangjin and Gyeongsang National University strains. The lowest variance (8.87 %) was between the Jangsu and Tongyeong strains. In addition, the genetic distance between the populations showed that Jangsu and Tongyeong formed one branch (they were very similar genetically). The Dangjin and the Gyeongsang National University strains appeared to form a second branch. Furthermore, the crossbred formed one branch with the Dangjin and the Gyeongsang National University strains. Therefore, the results of this study can be used as basic data to reduce unnecessary inbreeding and genetic resource flow between the KNBG populations. The basic data indicates the uniqueness of the genetic resources of the domestic lineage. These findings provide a basis for differentiating KNBG and Crossbred goats to use to improve the desirable characteristics of this species.

A Case of Posttransplant Lymphoproliferative Disease(PTLD) Following Renal Transplantation in a Child (소아에서 신이식후 발생한 Posttransplant Lymphoproliferative Disease(PTLD) 1례)

  • Eun, Byung-Wook;Park, Eun-Sil;Lee, Seong-Yong;Hahn, Hye-Won;Jang, Ju-Young;Park, Kyung-Mi;Kim, Chul-Woo;Kang, Gin-Han;Ko, Jae-Seong;Ha, Il-Soo;Lee, Hoan-Jong;Cheong, Hae-Il;Seo, Jeong-Kee
    • Childhood Kidney Diseases
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    • v.6 no.1
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    • pp.123-130
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    • 2002
  • Posttransplant lymphoproliferative disease (PTLD) represents a diverse lymphoproliferative disorder ranging front nonspecific reactive hyperplasia to malignant immunoblastic sarcoma developed in a setting of immunosuppression following organ or cellular transplantation. It is often associated with Epstein-Barr virus (EBV) infection and high dose immunosuppression. PTLD after renal transplantation was reported at first in adult in Korea in 1997. In children there have been several cases of PTLD after liver transplantation but PTLD after renal transplantation has not been reported. This is a case report of PTLD developed 4 months after renal transplantation in a 9-year-old boy. The major clinical manifestations were fever, multiple lymph nodes enlargement and blood-tinged stool. EBV was detected by in-situ hybridization in the enlarged cervical lymph node and the colonic tissue. Histological examination revealed B-cell lineage. Use of ganciclovir and reduction of the immunosuppression level resulted in complete remission of PTLD. This is the first pediatric case report of PTLD following renal transplantation in Korea. (J Korean Soc Pediatr Nephrol 2002 ; 6 : 123-30)

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Establishment of Functional Cells for Vascular Defect Disease from Human Embryonic Stem Cell via Region Sorting Depending on Cell Volume (세포 크기 차이를 이용한 유세포 분석을 통한 인간배아줄기세포 유래 기능성 혈관세포의 확립)

  • Lee, Ji-Hye;Kim, Ju-Mi;Chung, Hyung-Min;Chae, Jung-Il
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.364-373
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    • 2011
  • Human embryonic stem cells have been highlighted as a valuable cellular source in the regenerative medicine field, due to their pluripotency. However, there is the challenge of the establishment of specific functional cell type forms of undifferentiated human embryonic stem cells (hESC). To establish and purify functional cell types from hESCs, we differentiated undifferentiated hESCs into vascular lineage cells and sorted the specific cell population from the whole cell population, depending on their cell volume, and compared them with the non-sorted cell population. We observed that about 10% of the PECAM positive population existed in the VEGF induced differentiating human embryoid body (hEB), and differentiated hEBs were made into single cells for cell transplantation. After making single cells, we performed cell sorting using a fluorescence-activated cell sorter (FACs), according to their cell volume on the basis of FSC region gating, and compared their therapeutic capacity with the non-sorted cell population through cell transplantation into hindlimb ischemic disease model mice. 4 Weeks after cell transplantation, the recovery rate of blood perfusion reached 54% and 17% in the FSC regions of sorted cells- and non-sorted cells, respectively. This result suggests that derivation of a functional cell population from hESCs can be performed through cell sorting on the basis of cell volume after preliminary differentiation induction. This approach may then greatly contribute to overcoming the limitations of marker sorting.

Differential Potential of Stem Cells Following Their Origin - Subacromial Bursa, Bone Marrow, Umbilical Cord Blood - (줄기세포의 분화능의 기원에 따른 비교 - 견봉하 점액낭, 골수, 탯줄 혈액 -)

  • Sim, Sung Woo;Moon, Young Lae;Kang, Jung Hun
    • Clinics in Shoulder and Elbow
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    • v.15 no.2
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    • pp.65-72
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    • 2012
  • Purpose: To evaluate the differentiation potential of stem cells and their immunophenotype from 3 different sources. Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord blood. We obtained the subacromial bursal tissue and bone marrow from the patients undergoing shoulder surgery. After collecting the sample, we applied specific induction media for neurogenic, adipogenic and osteogenic differentiation. Also, flow-cytometry analysis was done to reveal the cell surface antigens. Results: We obtained 100% (8 cases) neural and adipogenic differentiation, but 62.5% (5 of 8 cases) osseous differentiation among the subacromial bursal tissue group. Bone marrow derived cells showed 100% neural (6 cases) and adipogenic (5 cases) differentiation, but 80% (4 of 5 cases) osseous differentiation. Umbilical cord blood derived cells revealed 97% (65 of 67 cases) neural, 53.7% (29 of 54 cases) adipogenic and 68.4% (39 of 57 cases) osseous differentiation. Immunophenotype analysis revealed that surface markers of bone marrow, subacromial bursal cell and umbilical cord blood derived mesenchymal stem cells are different from each other. Conclusions: Mesenchymal stem cells are potential agents in regenerative medicine and are characterized by expression of surface markers and by their differentiation potential. Our study with stem cells from subacromial bursal tissue, bone marrow and umbilical cord discovered that each stem cell has unique differentiation potential and function based on its origin. Various stem cells show multi-lineage differentiations in vitro which can be correlated to in vivo conditions.

Screening of Chicken Genes Related to Germ Cell Development (닭에서 생식세포 발달에 관여하는 유전자 검색)

  • Lee, Jee-Young;Kim, Hee-Bal;Kim, Duk-Kyung;Song, Ki-Duk;Lim, Jeong-Mook;Han, Jae-Yong
    • Journal of Animal Science and Technology
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    • v.49 no.2
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    • pp.183-194
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    • 2007
  • We examined the expression patterns of the chicken TCs(tentative consensus sequences) originated from GermOnline genes in various chicken tissues, applying information from GermOnline to chicken organisms. 42 TCs among 84 chicken homologous TCs from the pool of 84 genes related to germ cell lineage in mouse(10), rat(71) and human(3) had high homology based on a BLAST search. Of these, Hmgcs2 and Sycp3 was shown to be expressed in a testis- specific manner and a reproductive organ(testis and ovary)-specific manner, respectively, by RT- PCR analysis. Crmp4, Cyct, Ldhc, Epha7, Pcsk4 and Dnmt3a are expressed in brain, testis, and ovary. The characterization of chicken genes originated from GermOnline in this research may give an enormously useful source of information related to germ cell development.

Natural hybridization of Iris species in Mt. Palgong-san, Korea (팔공산 금붓꽃 계열의 자연 잡종 현상)

  • Son, OGyeong;Son, Sung-Won;Suh, Gang-Uk;Park, SeonJoo
    • Korean Journal of Plant Taxonomy
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    • v.45 no.3
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    • pp.243-253
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    • 2015
  • Series Chinensis, Genus Iris, endemic to the far regions of East Asia, consists of four species and related varieties. This series is divided into two major groups (I. rossii and I. minutiaurea complex). In this study, the ITS region and matK gene sequences within nuclear ribosomal DNA and plastid DNA were analyzed in order to investigate the phylogenetic relationships among the I. minutiaurea complex (I. minutiaurea, I. odaesanensis, and I. koreana) and the taxonomic identities of a putative hybrid in Mt. Palgong. In the internal transcribed spacer (ITS1, 5.8S, and ITS2) region, a total of 106 cloned genomic sequences from three taxa were obtained to study the intragenomic polymorphisms of the ITS regions. Three taxa revealed high levels of intragenomic polymorphisms, indicative of incomplete nrDNA concerted evolution. This incomplete ITS concerted evolution in the series Chinensis may be linked to the recent species divergence and frequent interspecies hybridization of the series Chinensis. In the matK gene, three taxa were fairly separated by eleven variable sites. In eight individuals collected on Mt. Palgong, putative hybrids between I. odaesanensis and I. minutiaurea were clustered in the I. minutiaurea clade in the NJ (neighbor-joining) tree based on the matK gene. However, in the ITS tree, some of them were clustered in the I. odaesanensis clade and others were clustered in the I. minutiaurea clade. Therefore, the individuals on Mt. Palgong were formed by the hybridization between two taxa (I. odaesanensis and I. minutiaurea) and not through the lineage of I. koreana.

Isolation of human mesenchymal stem cells from the skin and their neurogenic differentiation in vitro

  • Byun, Jun-Ho;Kang, Eun-Ju;Park, Seong-Cheol;Kang, Dong-Ho;Choi, Mun-Jeong;Rho, Gyu-Jin;Park, Bong-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.38 no.6
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    • pp.343-353
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    • 2012
  • Objectives: This aim of this study was to effectively isolate mesenchymal stem cells (hSMSCs) from human submandibular skin tissues (termed hSMSCs) and evaluate their characteristics. These hSMSCs were then chemically induced to the neuronal lineage and analyzed for their neurogenic characteristics in vitro. Materials and Methods: Submandibular skin tissues were harvested from four adult patients and cultured in stem cell media. Isolated hSMSCs were evaluated for their multipotency and other stem cell characteristics. These cells were differentiated into neuronal cells with a chemical induction protocol. During the neuronal induction of hSMSCs, morphological changes and the expression of neuron-specific proteins (by fluorescence-activated cell sorting [FACS]) were evaluated. Results: The hSMSCs showed plate-adherence, fibroblast-like growth, expression of the stem-cell transcription factors Oct 4 and Nanog, and positive staining for mesenchymal stem cell (MSC) marker proteins (CD29, CD44, CD90, CD105, and vimentin) and a neural precursor marker (nestin). Moreover, the hSMSCs in this study were successfully differentiated into multiple mesenchymal lineages, including osteocytes, adipocytes, and chondrocytes. Neuron-like cell morphology and various neural markers were highly visible six hours after the neuronal induction of hSMSCs, but their neuron-like characteristics disappeared over time (24-48 hrs). Interestingly, when the chemical induction medium was changed to Dulbecco's Modified Eagle Medium (DMEM) supplemented with fetal bovine serum (FBS), the differentiated cells returned to their hSMSC morphology, and their cell number increased. These results indicate that chemically induced neuron-like cells should not be considered true nerve cells. Conclusion: Isolated hSMSCs have MSC characteristics and express a neural precursor marker, suggesting that human skin is a source of stem cells. However, the in vitro chemical neuronal induction of hSMSC does not produce long-lasting nerve cells and more studies are required before their use in nerve-tissue transplants.

Effect of Dendritic Cell Based Cancer Vaccine Using Allogeneic Tumor Cell Lysate in Melanoma Pulmonary Metastasis Model (동종 종양 세포 용해액을 이용한 수지상 세포 항암 백신의 흑색종 폐암 전이 모델에서의 효과 연구)

  • Lee, Young-Joon;Kim, Myung-Joo;In, So-Hee;Choi, Ok-Mi;Baek, So-Young;Kwon, Young-Do;Lee, Hyun-Ah
    • IMMUNE NETWORK
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    • v.5 no.3
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    • pp.163-171
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    • 2005
  • Background: To perform the successful dendritic cell-based cancer immunotherapy one of the main issues to be solved is the source of antigen for DC pulsing. Limitations occur by using auto-tumor lysate due to the difficulties obtaining enough tumor tissue(s) quantitatively as well as qualitatively. In this study the possibility of allogeneic tumor cell lysate as a DC pulsing antigen has been tested in mouse melanoma pulmonary me tastasis model. Methods: B16F10 melanoma cells $(1{\timeS}10^5/mouse)$ were inoculated intra venously into the C57BL/6 mouse. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 (1,000 U/ml each) for 7 days and pulsed with lysate of either autologous B16F10 (B-DC), allogeneic K1735 (C3H/He origin; K-DC) or CloneM3 (DBA2 origin; C-DC) melanoma cells for 18 hrs. Pulsed-DCs $(1{\times}10^6/mouse)_{[CGP1]}$ were injected i.p. twice with one week interval starting from the day 1 after tumor cell inoculation. Results: Without observable toxicity, allogeneic tumor cell lysate pulsed-DC induced the significantly better anti-tumor response (tumor scale: $2.7{\pm}0.3,\;0.7{\pm}0.3\;and\;0.3{\pm}0.2$ for saline, B-DC and C-DC treated group, respectively). Along with increased tumor specific lymphocyte proliferations, induction of IFN-${\gamma}$ secretion against both auto- and allo-tumor cell lysates was observed from the DC treated mice. (w/B16F10-lysate: $44.97{\pm}10.31,\;1787.94{\pm}131.18,\;1257.15{\pm}48.27$, w/CloneM3 lysate: 0, $1591.13{\pm}1.83,\;1460.47{\pm}86.05pg/ml$ for saline, B-DC and C-DC treated group, respectively) Natural killer cell activity was also increased in the mice treated with tumor cell lysate pulsed-DC ($8.9{\pm}_{[CGP2]}0.1,\;11.6{\pm}0.8\;and\;12.6{\pm}0.7%$ specific NK activity for saline, B-DC and C-DC treated group, respectively). Conclusion: Conclusively, promising data were obtained that allogeneic-tumor cell lysate can be used as a tumor antigen for DC-based cancer immunotherapy.

Evolutionary Relationship of Liobagrus mediadiposalis (Teleostei: Amblycipitidae) Populations in Korea Inferred from Cytochrome b DNA Sequences (한국 고유종인 자가사리(Liobagrus mediadiposalis) 지역개체군의 분자진화적 유연관계)

  • Kim, Maeng Jin;Han, Song-Hun;Yang, Hye-Young;Jo, Mi-Ran;Chung, Sang-Chul;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.18 no.4
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    • pp.329-338
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    • 2006
  • Phylogenetic relationships and DNA polymorphism among local populations of the Korean native catfish species, Liobagrus mediadiposalis, have been investigated based on mitochondrial cytochrome b DNA sequences. As a result, three genetically distinct groups of local populations were recognized based on the phylogenetic tree constructed. The first group was called "Nakdong-river group" that included the local populations from Geumho-river, Gyeongho-river, and Deokcheon-river; the second one was "Geum-river group"; the third one was represented as "Seomjin-river group" that included the samples from Seomjin-river, Dongjin-river and Geokum-do. The phylogeny also implied that the ancestral group of L. mediadiposalis have first evolved to Nakdong-river group, and later two local populations (Geum-river and Seomjinriver group) were diverged from the other lineage. DNA polymorphisms we observed were 4.4~4.7% between Geum-river group and Seomjin-river group, 5.1~5.5% between Seomjinriver group and Nakdong-river group, and 5.5~5.7% between Nakdong-river group and Geumriver group. These results indicated the long period of geographic isolation due to the river system in Korea caused such high degrees of DNA polymorphisms between local populations of L. mediadiposalis.