• Title/Summary/Keyword: Cord blood transplantation

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Proliferation, Apoptosis, and Telomerase Activity in Human Cord Blood CD34+ Cells Cultured with Combinations of Various Cytokines

  • Ahn, Myung-Ju;Lee, Hye-Sook;Jang, Mi-Yune;Choi, Jung-Hye;Lee, Young-Yeul;Park, Hyung-Bae;Lee, Yong-Sung
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.422-428
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    • 2003
  • Umbilical cord blood (UCB), a rich source of hematopoietic stem/progenitor cells, has been proposed as an alternative to bone marrow and peripheral blood for transplantation treatment. Ex vivo expansion of cord blood stem cells could make the use of cord blood transplant feasible even for adult patients. However, the optimal cytokine cocktail for expansion of stem cells is yet to be established. This study compares proliferation, apoptosis, and telomerase activities in human cord blood stem cells cultured ex vivo with FLT3 ligand (FL)/thrombopoietin (TPO) or FL/TPO/stem cell factor (SCF), with a view to determine optimal combination of cytokines. CD34+ cells were cultured in DMEM containing either FL (50 ng/ml) and TPO (10 ng/ml) (FT group) or FL (50 ng/ml), TPO (10 ng/ml) and SCF (50 ng/ml) (FTS group). The cell proliferation rate was ten times higher in the FTS group. Although cells cultured with the two different combinations of cytokines were maintained for a long term (up to 8 weeks), a large number of cells underwent differentiation during this period. Cells cultured in FTS displayed lower levels of apoptosis compared to those of the FT group during the Initial 7 days of culture. The CD34+ fraction in both groups was markedly decreased to $21-30\%$ , and only $5-6\%$ was detected at 14 days of culture. Telomerase activity detected in human CD34+ cord blood at low levels was upregulated during the early phase of culture and decreased to baseline levels in the later phase. The telomerase activity of cord blood cultured in FT was lower than that of the FTS group. Our results suggest that, on adding stem cell factors to the FT cytokines, cultured CD34+ cord blood cells display a greater degree of cell proliferation and decreased apoptosis. However, during CD34+ cord blood cell culture, a Barge number of cells undergo differentiation, indicating that more potent novel cytokines or new culture conditioning methods should be developed to maintain their ability to engraft and sustain long-term hematopoiesis.

Expression of HLA and Mixed Lymphocyte Reaction of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood (제대혈 유래 중간엽줄기세포에서 HLA의 발현과 Mixed Lymphocyte Reaction)

  • Lee, Hyo-Jong;kang, Sun-Young;Park, Se-Jin;Lee, Seung-Yong;Lee, Hee-Chun;Koh, Phil-Ok;Park, Ji-Kwon;Paik, Won-Young;Yeon, Seong-Chan
    • Journal of Veterinary Clinics
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    • v.28 no.4
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    • pp.399-402
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    • 2011
  • In recent years, the mesenchymal stem cells (MSC) derived from various tissues have been widely tested for developing cell therapies, tissue repair and transplantation. Although there has been much interest in the immunomodulatory properties of MSC and their immunologic reactions following autologous, allogeneic and xenogenic transplantation of MSC in vivo, up to date, the expression of immunogenic markers, such as class I and II human leukocyte antigens (HLA), after differentiation of human umbilical cord blood (hUCB)-derived MSC has been poorly investigated and require extensive in vitro and in vivo testing. In this experiment, the expression of the HLA-ABC and HLA-DR on hUCB-derived MSC have been tested by immunocytochemical staining. The undifferentiated MSC were moderately stained for HLA-ABC but very weakly for HLA-DR. In order to investigate the inhibitory effect of allogeneic lymphocytes on proliferation of MSC, the MSC were cultured in the presence or absence of peripheral allogeneic lymphocytes stimulated with concanavalin A. The allogeneic lymphocytes did not significantly inhibit MSC proliferation. We conclude that hUCB-MSC expressed moderately class I HLA antigen while almost negatively class II HLA antigen. The MSC have an immunomodulatory effect which can suppress the allogeneic response of lymphocytes. These in vitro data suggest that allogeneic MSC derived from cord blood can be useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.

A Successful Transplant of HLA-3 Loci Mismatched Umbilical Cord Blood into a Patient with Acute Myeloid Leukemia from an Unrelated Donor (급성 골수구성 백혈병 환아에서 비혈연간 주조직 적합 항원 3부위 불일치 제대혈 조혈모세포 이식 성공 1례)

  • Kwon, Byoung Chul;Shin, Kyung Mi;Won, Sung Chul;Lyu, Chuhl Joo;Yang, Chang Hyun;Kim, Hyun Ok
    • Clinical and Experimental Pediatrics
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    • v.45 no.7
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    • pp.912-916
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    • 2002
  • Cord blood is a useful source of allogeneic hematopoietic stem cells for bone marrow reconstitution. The number of umbilical cord blood transplants is increasing worldwide. In this a case 15-month-old boy with acute myeloid leukemia was treated with umbilical cord blood transplant from an HLA-3 loci mismatched unrelated donor. Granulocyte recovery greater than $500/mm^3$ occurred at day 49, and the platelet recovered greater than $20,000/mm^3$ independent of transfusion at day 81 after stem cell infusion.

ENDOTHELIAL PROGENITOR CELLS AND MESENCHYMAL STEM CELLS FROM HUMAN CORD BLOOD (제대혈 내피기원세포 및 간엽줄기세포의 분화에 대한 연구)

  • Kim, Eun-Seok;Kim, Hyun-Ok
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.1
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    • pp.39-45
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    • 2005
  • Stem cell therapy using mesenchymal stem cells(MSCs) transplantation have been paid attention because of their powerful proliferation and pluripotent differentiating ability. Although umbilical cord blood (UCB) is well known to be a rich source of hematopoietic stem cells with practical and ethical advantages, the presence of mesenchymal stem cells (MSCs) in UCB has been controversial and it remains to be validated. In this study, we examine the presence of MSCs in UCB harvests and the prevalence of them is compared to that of endothelial progenitor cells. For this, CD34+ and CD34- cells were isolated and cultured under the endothelial cell growth medium and mesenchymal stem cell growth medium respectively. The present study showed that ESC-like cells could be isolated and expanded from preterm UCBs but were not acquired efficiently from full-terms. They expressed CD14-, CD34-, CD45-, CD29+, CD44+, CD105+ cell surface marker and could differentiate into adipogenic and osteogenic lineages. Our results suggest that MSCs are fewer in full-term UCB compared to endothelial progenitor cells.

Estrogen Mediates Ischemic Damage and the Migration of Human Umbilical Cord Blood Cells

  • Kim, Jee-Yun;Yu, Seong-Jin;Kim, Do-Rim;Youm, Mi-Young;Lee, Chae-Kwan;Kang, Sung-Goo
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.71-71
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    • 2003
  • Human umbilical cord blood cells(HUCBC) are rich in mesenchymal progenitor cells, endothelial cell precursors and hematopoietic cells. HUCBC have been used as a source of transplantable stem and progenitor cells. However, little is known about survival and development of HUCBC transplantation in the CNS. Estrogen has a neuroprotective potential against oxidative stress-induced cell death so has an effect on reducing infarct size of ischemic brain. We investigated the potential use of HUCBC as donor cells and tested whether estrogen mediates intravenously infused HUCBC enter and survive in ischemic brain. PKH26 labeled mononuclear fraction of HUCBC were injected into the tail vein of ischemic OVX rat brain with or without $17\beta$-estradiol valerate(EV). Under fluorescence microscopy, labeled cells were observed in the brain section. Significantly more cells were found in the ischemic brain than in the non-ischemic brain. HUCBC transplanted into ischemic brain could migrate and survive. Some of cells have shown neuronal like cells in hippocampus, striatum and cortex tissues. These result suggest that estrogen reduces ischemic damage and increases the migration of human umbilical cord blood cells. This Study was supported by the Korea Science and Engineering Foundation(KOSEF) though the Biohealth Products Research Center(BPRC), Inje University, Korea.

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Nanosphere Form of Curcumin Stimulates the Migration of Human Umbilical Cord Blood Derived Mesenchymal Stem Cells

  • Kim, Do-Wan;Kim, Ju Ha;Lee, Sei-Jung
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2020.10a
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    • pp.221-221
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    • 2020
  • Curcumin, a hydrophobic polyphenol derived from turmeric, has been used a food additive and as a herbal medicine for the treatment of various diseases. In the present study, we found the functional role of a nanosphere loaded with curcumin (CN) in the promotion of the motility of human umbilical cord blood derived mesenchymal stem cells (hUCB-MSCs) during the wound closure. We found that the efficacy of hUCB-MSCs migration induced by CN was 1000-fold higher than that of curcumin powder. CN significantly increased the motility of hUCB-MSCs by activating c-Src, which is responsible for the phosphorylation of protein kinase C (PKC) and extracellular signal-regulated kinase (ERK). CN induced the expression levels of α-actinin-1, profilin-1 and filamentous-actin, as regulated by the phosphorylation of nuclear factor-kappa B during its promotion of cell migration. In a mouse skin excisional wound model, we found that transplantation of UCB-MSCs pre-treated with CN enhances wound closure, granulation, and re-epithelialization at mouse skin wound sites. These results indicate that CN is a functional agent that promotes the mobilization of UCB-MSCs for cutaneous wound repair.

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Ex vivo Expansion and Clonal Maintenance of CD34+ Selected Cells from Cord Blood and Peripheral Blood (제대혈 및 말포혈로부터 분리한 CD34 양성 세포의 체외 증폭 및 클론 유지)

  • Kim, Soon Ki;Ghil, Hye Yoon;Song, Sun U.;Choi, Jong Weon;Park, Sang Kyu
    • Clinical and Experimental Pediatrics
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    • v.48 no.8
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    • pp.894-900
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    • 2005
  • Purpose : Because of the unavailability of marrow transplantation, umbilical cord blood (CB) is increasingly being used. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from cord blood source and mobilized peripheral blood (PB) in a serum-free media. Methods : The CD34+ cells, selected from CB and mobilized PB, were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at culture days 0, day 4, day 7, and day 14 with various growth factors. Results : The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the PB at day 7 (2 fold increase than PB). The CB-selected CD34+ cells produced more BFU-E colonies than did the PB on culture at days 7 and at day 14. Also, the CB-selected CD34+ cells produced more CFU-Mk colonies than did the PB on culture at day 4 and at day 7. Conclusion : The ex vivo expansion of the CB cells may be promising in producing total cellular expansion, CFU-Mk and BFU-E compared with PB for 7 to 14 days. The growth factors combination including megakaryocyte growth and development, flt3-ligand and interleukin-3 showed more expansion in the view of total cells and clonal maintenance compared with less combination.

Blood Vessel Regeneration using Human Umbilical Cord-derived Endothelial Progenitor Cells in Cyclophosphamide-treated Immune-deficient Mice

  • Kwon, Soon-Keun;Ko, Yu-Jin;Cho, Tae-Jun;Park, Eu-Gene;Kang, Byung-Chul;Lee, Gene;Cho, Jae-Jin
    • International Journal of Oral Biology
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    • v.36 no.3
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    • pp.117-122
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    • 2011
  • Endothelial cells are a vital constituent of most mammalian organs and are required to maintain the integrity of these tissues. These cells also play a major role in angiogenesis, inflammatory reactions, and in the regulation of thrombosis. Angiogenesis facilitates pulp formation and produces the vessels which are essential for the maintenance of tooth homeostasis. These vessels can also be used in bone and tissue regeneration, and in surgical procedures to place implants or to remove cancerous tissue. Furthermore, endothelial cell regeneration is the most critical component of the tooth generation process. The aim of the present study was to stimulate endothelial regeneration at a site of acute cyclophosphamide (CP)-induced endothelial injury by treatment with human umbilical cord-derived endothelial/mesenchymal stem cells (hEPCs). We randomly assigned 16 to 20-week-old female NOD/SCID mice into three separate groups, a hEPC ($1{\times}10^5$ cells) transplanted, 300mg/kg CP treated and saline (control) group. The mice were sacrificed on days 5 and 10 and blood was collected via the abdominal aorta for analysis. The alanine transaminase (ALT), aspartate aminotransferase (AST), serum alkaline phosphatase (s-ALP), and albumin (ALB) levels were then evaluated. Tissue sections from the livers and kidneys were stained with hematoxylin and eosin (HE) for microscopic analysis and were subjected to immunohistochemistry to evaluate any changes in the endothelial layer. CP treatment caused a weight reduction after one day. The kidney/body weight ratio increased in the hEPC treated animals compared with the CP only group at 10 days. Moreover, hEPC treatment resulted in reduced s-ALP, AST, ALT levels compared with the CP only group at 10 days. The CP only animals further showed endothelial injuries at five days which were recovered by hEPC treatment at 10 days. The number of CD31-positive cells was increased by hEPC treatment at both 5 and 10 days. In conclusion, the CP-induced disruption of endothelial cells is recovered by hEPC treatment, indicating that hEPC transplantation has potential benefits in the treatment of endothelial damage.

Aplastic anemia (재생불량빈혈(Aplastic anemia))

  • Kim, Hack Ki
    • Clinical and Experimental Pediatrics
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    • v.50 no.6
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    • pp.519-523
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    • 2007
  • Aplastic anemia is a rare disease, which is characterized by pancytopenia and hypocellular bone marrow without infiltration of abnormal cells or fibrosis. The incidence in Asia is higher than in the West and new cases are diagnosed at a rate of 5.1 per million pediatric populations per year in Korea. The pathophysiology is understood roughly by defective hematopoiesis, impaired bone marrow micro-environment and immune mechanism. Treatments are performed on basis of pathogenesis and selected depending on the severity. Immunosuppressive therapy with antilymphocyte or antithymocyte globulin and cyclosporine is effective in the majority of patients but has some problems including relapse or clonal evolution. Recently, there have been clinical trials of immunosuppression with hematopoietic growth factors or other drugs. Allogeneic hematopoietic stem cell transplantation (HSCT) is curative in children with severe aplastic anemia. The overall survival in HSCT from HLA-identical sibling is higher than alternative donor, including HLA matched unrelated donor or cord blood. We have to consider quality of life after HSCT because of high survival rate. However, chronic graft versus host disease and graft failure are important factors that affect the quality of life and overall survival. We need further investigation to make new regimens aimed at overcoming these risk factors and perform clinical trials.