• Title/Summary/Keyword: Hematopoietic stem cells

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Toxicity and Biomedical Imaging of Fluorescence-Conjugated Nanoparticles in Hematopoietic Progenitor Cells

  • Min, Gye-Sik;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.503-510
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    • 2011
  • Cellular uptake of nanoparticles for stem cell labeling and tracking is a critical technique for biomedical therapeutic applications. However, current techniques suffer from low intracellular labeling efficiency and cytotoxic effects, which has led to great interest in the development of a new labeling strategy. Using silica-coated nanoparticles conjugated with rhodamine B isothiocyanate (RITC) (SR), we tested the cellular uptake efficiency, biocompatibility, proliferation or differentiation ability with murine bone marrow derived hematopoietic stem/progenitor cells. The bone marrow hematopoietic cells showed efficient uptake with SR with dose or time dependent manner and also provided a higher uptake on hematopoietic stem/progenitor cells. Biocompatibility tests revealed that the SR had no deleterious effects on cell cytotoxicity, proliferation, or multi-differentiation capacities in vitro and in vivo. SR nanoparticles are advantageous over traditional labeling techniques as they possess a high level of cellular internalization without limiting the biofunctionality of the cells. Therefore, SR provides a useful alternative for gene or drug delivery into hematopoietic stem/progenitor cells for basic research and clinical applications.

Expression and Characterization of Purinergic Receptor, $P2Y_{10}$ in Hematopoietic Stem Cells

  • Lee Eun-Jong;Kim Dong-Ku
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.109-115
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    • 2005
  • Hematopoietic stem cells (HSC) are multipotent cells that reside in the bone marrow and replenish all adult hematopoietic lineages throughoutthe lifetime. In this study, we analyzed the expression of receptors of $P2Y_{10}$, purinergic receptor families in murine hematopoietic stem cells, hematopoietic progenitor cells. In addition, the biological activity of $P2Y_{10}$ was investigated with B lymphocyte cell line, Ba/F3 in effect to cell growth and cell cycle. From the analysis of expression in hematopoieticstem cell. and progenitor with RT-PCR, $P2Y_{10}$ was strongly expressed in murine hematopoieticstem cells (c-kit+ Sca-l+ Lin-) and progenitor cell population, such as c-kit- Sca-l+ Lin-, c-kit+ Sca-l- Lin- and c-kit- Sca-l- Lin-. To investigate the biological effects by $P2Y_{10}$, retroviral vector from subcloned murine $P2Y_{10}$ cDNA was used fur gene introduction into Ba/F3 cells, and stable transfectant cells were obtained by flow cytometry sorting. In cell proliferation assay, the proliferation ability of $P2Y_{10}$ receptor gene­transfected cells was strongly inhibited, and the cell cycle was arrested at G1 phase. These result suggest that the $P2Y_{10}$ may be involved the biological activity in hematopoietic stem cells and immature B lymphocytes.

Forced Expression of HoxB4 Enhances Hematopoietic Differentiation by Human Embryonic Stem Cells

  • Lee, Gab Sang;Kim, Byung Soo;Sheih, Jae-hung;Moore, Malcolm AS
    • Molecules and Cells
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    • v.25 no.4
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    • pp.487-493
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    • 2008
  • HoxB4 has been shown to enhance hematopoietic engraftment by hematopoietic stem cells (HSC) from differentiating mouse embryonic stem cell (mESC) cultures. Here we examined the effect of ectopic expression of HoxB4 in differentiated human embryonic stem cells (hESCs). Stable HoxB4-expressing hESCs were established by lentiviral transduction, and the forced expression of HoxB4 did not affect stem cell features. HoxB4-expressing hESC-derived CD34+ cells generated higher numbers of erythroid and blast-like colonies than controls. The number of CD34+ cells increased but CD45+ and KDR+ cell numbers were not significantly affected. When the hESC derived CD34+ cells were transplanted into $NOD/SCID{\beta}2m-/-$ mice, the ectopic expression of HoxB4 did not alter their repopulating capacity. Our findings show that overexpression of HoxB4 in differentiating hESCs increases hematopoietic colony formation and hematopoietic cell formation in vitro, but does not affect in vivo repopulation in adult mice hosts.

The Intetions of University Students Regarding Donating Hematopoietic Stem Cells Based on the Theory of Planned Behavior (계획행위이론에 근거한 대학생의 조혈모세포 기증희망 등록의도)

  • Lim, Seungjoo;Cho, Sorin;Yang, Eunjung
    • Journal of muscle and joint health
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    • v.27 no.2
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    • pp.153-159
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    • 2020
  • Purpose: This study aims to identify the effect of university students' intention to donate hematopoietic stem cells based on the theory of planned behavior. Methods: The subjects include university students who visited the campaign for the registration of hematopoietic stem cell donation held at H university on September 28, 2019 and October 2, 2019. Results: The intention to register for hematopoietic stem cell donation and empirical attitude (r=.72, p<.001), instrumental attitude (r=.64, p<.001), directive norm (r=.53, p<.001), technical norms (r=.55, p<.001) and self-efficacy (r=.86, p<.001) showed a significant positive correlation. The multiple regression analysis revealed that self-efficacy (β=.66) and empirical attitude (β=.23) were the most influential factors. Conclusion: Educational and promotional programs to increase the intention to register for hematopoietic stem cell donation need to be developed to help increase students' self-efficacy and help them develop a positive experiential attitude. In addition, further research is needed to determine whether the intention to register in hematopoietic stem cell donation among university students can lead to actual registration and donation after registration.

Development of Natural Killer Cells from Hematopoietic Stem Cells

  • Yoon, Suk Ran;Chung, Jin Woong;Choi, Inpyo
    • Molecules and Cells
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    • v.24 no.1
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    • pp.1-8
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    • 2007
  • Natural killer (NK) cells play a crucial role in innate immune system and tumor surveillance. NK cells are derived from $CD34^+$hematopoietic stem cells and undergo differentiation via precursor NK cells in bone marrow (BM) through sequential acquisition of functional surface receptors. During differentiation of NK cells, many factors are involved including cytokines, membrane factors and transcription factors as well as microenvironment of BM. NK cells express their own repertoire of receptors including activating and inhibitory receptors that bind to major histocompatibility complex (MHC) class I or class I-related molecules. The balance between activating and inhibitory receptors determines the function of NK cells to kill targets. Binding of NK cell inhibitory receptors to their MHC class I-ligand renders the target cells to be protected from NK cell-mediated cytotoxicity. Thus, NK cells are able to discriminate self from non-self through MHC class I-binding inhibitory receptor. Using intrinsic properties of NK cells, NK cells are emerging to apply as therapeutic agents against many types of cancers. Recently, NK cell alloactivity has also been exploited in killer cell immunoglobulin-like receptor mismatched haploidentical stem cell transplantation to reduce the rate of relapse and graft versus host disease. In this review, we discuss the basic mechanisms of NK cell differentiation, diversity of NK cell receptors, and clinical applications of NK cells for anti-cancer immunotherapy.

Profiling of Differentially Expressed Genes in Human Stem Cells by cDNA Microarray

  • Kim, Chul Geun;Lee, Jong Joo;Jung, Dae Young;Jeon, Jinseon;Heo, Hyen Seok;Kang, Ho Chul;Shin, June Ho;Cho, Yoon Shin;Cha, Kyung Joon;Kim, Chan Gil;Do, Byung-Rok;Kim, Kyung Suk;Kim, Hyun-Soo
    • Molecules and Cells
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    • v.21 no.3
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    • pp.343-355
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    • 2006
  • Stem cells are unique cell populations with the ability to undergo both self-renewal and differentiation, although a wide variety of adult stem cells as well as embryonic stem cells have been identified and stem cell plasticity has recently been reported. To identify genes implicated in the control of the stem cell state as well as the characteristics of each stem cell line, we analyzed the expression profiles of genes in human embryonic, hematopoietic ($CD34^+$ and $CD133^+$), and mesenchymal stem cells using cDNA microarrays, and identified genes that were differentially expressed in specific stem cell populations. In particular we were able to identify potential hESC signature-like genes that encode transcription factors (TFAP2C and MYCN), an RNA binding protein (IMP-3), and a functionally uncharacterized protein (MAGEA4). The overlapping sets of 22 up-regulated and 141 down-regulated genes identified in this study of three human stem cell types may also provide insight into the developmental mechanisms common to all human stem cells. Furthermore, our comprehensive analyses of gene expression profiles in various adult stem cells may help to identify the genetic pathways involved in self-renewal as well as in multi-lineage specific differentiation.

Efficient Gene Delivery into Hematopoietic Stem Cells by Intra-Bone Marrow Injection of Retrovirus (IBM 이식을 통한 골수 조혈 줄기 세포에의 효과적인 유전자 도입)

  • Lee, Byun-Joo;Lee, Yong-Soo;Kim, Hye-Sun;Kim, Yu-Kyung;Kim, Jae-Hwan;Park, Jin-Ki;Chung, Hak-Jae;Chang, Won-Kyong;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.9-14
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    • 2008
  • Efficient gene transfer into hematopoietic stem cells is a great tool for gene therapy of hematopoietic disease. Retrovirus have been extensively used for gene delivery and gene therapy. However, current in vitro gene transfer has some obstacles suck as induction of differentiation loss of self-renewal capacity, and down-regulation of homing efficiency for in vitro hematopoietic stem cells transplantation. To overcome these problems, we developed efficient in vitro retroviral transfer technique by direct intra-bone marrow injection (IBM). We identified effective retrovirus gene transfer in bone marrow hematopoietic cells in vitro. Two weeks after retrovirus transfer via IBM injection, we observed stable EGFP gene expression in bone marrow, lymph node, spleen, and liver cells. In addition, $6.4{\pm}2.7%$ of hematopoietic stem/progenitor cells were expressed EGFP transgene from flow cytometry analysis. Our results demonstrate that in vitro retrovirus gene transfer via IBM injection can provide a viable alternative to current or moo gene transfer approach.

Mesenchymal stem cells and osteogenesis

  • Jung, Cho-Rok;Kiran, Kondabagil R.;Kwon, Byoung S.
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.179-186
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    • 2001
  • Bone marrow stroma is a complex tissue encompassing a number of cell types and supports hematopiesis, differentiation of erythreid, nyel and lymphoid lineages, and also maintains undifferentiated hematopoietic stem cells. Marrow-derived stem cells were composed of two populations, namely, hematopoietic stem cells that can differentiate into blood elements and mesenchymal stem cells that can give rise to connective tissues such as bone, cartilage, muscle, tendon, adipose and stroma. Differentiation requires environmental factors and unique intracellular signaling. For example, $TGF-{\beta}$ or BMP2 induces osteoblastic differentiation of mesenchymal stem are very exciting. However, the intrinsic controls involved in differentiation of stem cells are yet to be understood properly in order to exploit the same. This review presents an overview of the recent developments made in mesenchymal stem cell research with respect to osteogenesis.

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The Effects of Extracts Mixture Drink from Inonotus Obliquus, Phellinus Linteus and Ganoderma Lucidum on Hematopoietic Stem Cells and Lymphocyte Subset of Blood in Human (차가버섯, 상황버섯 및 영지버섯 복합추출물 복용이 인체의 혈중 조혈모세포와 면역세포에 미치는 영향)

  • Bae, Hyung-Suk;Kang, Sung-Keun;Shin, Il-Seob;Woo, Sang-Kyu;Kim, Yun-Joung;Kim, Mi-Ae;Ra, Jeong-Chan
    • Journal of Food Hygiene and Safety
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    • v.24 no.1
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    • pp.78-85
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    • 2009
  • This study was performed to investigate the effect of extract mixture(IPGE) drink from Inonotus Obliquus, Phellinus Linteus and Ganoderma Lucidum on hematopoietic stem cells and lymphocyte subset[lymphocyte, $CD4^+T$ cell, $CD8^+T$ cell, Natural Killer(NK) cells] of blood in 37 participants who were healthy and about $40{\sim}70$ years old. They were divided into two groups; extract mixture drink administration group(n=27) and placebo administration group(n=12). They were given the test drink daily for 4 weeks. Blood was obtained from the subjects every two week in the beginning of administration day to evaluate the $CD34^+$ hematopoietic stem cells and immune cells. As results, $CD34^+$ hematopoietic stem cells were significantly increased after taking IPGE drink for 4 weeks compared to that before taking the drink (p<0.001). There was no significant changes in number of lymphocytes, $CD4^+T$ cells, $CD8^+T$ cells, NK cells and in the ratio of $CD4^+/CD8^+$ cell after taking the test drink. From these results, it was suggested that IPGE have a good health effect by promoting the proliferation of the hematopoietic stem cells.

Effective Reconstitution of Porcine Hematopoietic Cells in Newborn NOD/SCID Mice Xenograft (돼지 골수 조혈 세포의 이종 마우스 동물 모델 생체 증식 및 분화 특성)

  • Lee, Yong-Soo;Lee, Hyun-Joo;Kim, Tea-Sik;Kim, Hye-Sun;Kim, Yoo-Kyong;Kim, Jae-Hwan;Park, Jin-Ki;Chung, Hak-Jae;Chang, Won-Kyong;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.1-7
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    • 2008
  • The SCID-repopulation cells(SRCs) assay has been widely used to determine the self-renewal capacity of hematopoietic stem cells (HSCs). In this study, we tested the repopulating efficiency of porcine bone marrow derived hematopoietic stem cells using nonobese diabetic/severe combined immunodieficient (NOD/SCID) mice which was inherited immunodeficiency mire with defect of T cells, B cells, and low activity of NK cells. We transplanted porcine bone marrow hematopoietic stem/progenitor cells with intraperitoneal injection into neonate NOD/SCID mice. We confirmed efficient reconstitution activity of inoculated porcine hematopoietis cells in variety of organs of NOD/SCID mice. Interestingly, pig $CD3^+$ T lymphocytes detected with high level in liver($15.6{\pm}3.7%$), spleen($5.6{\pm}3.0%$), thymus($1.5{\pm}1.3%$), and BM($2.3{\pm}0.9%$), respectively. These data imply that microenvironment of neonate NOD/SCID mice is very efficient for proliferation and differentiation of porcine T cells, and can be useful for the study of T cells development and renogeneic organ transplantation.