• 제목/요약/키워드: pluripotent

검색결과 210건 처리시간 0.032초

Stem Cell Biology, 최근의 진보 (Recent Advancement in the Stem Cell Biology)

  • 한창열
    • Journal of Plant Biotechnology
    • /
    • 제33권3호
    • /
    • pp.195-207
    • /
    • 2006
  • Stem cells are the primordial, initial cells which usually divide asymmetrically giving rise to on the one hand self-renewals and on the other hand progenitor cells with potential for differentiation. Zygote (fertilized egg), with totipotency, deserves the top-ranking stem cell - he totipotent stem cell (TSC). Both the ICM (inner cell mass) taken from the 6 days-old human blastocyst and ESC (embryonic stem cell) derived from the in vitro cultured ICM have slightly less potency for differentiation than the zygote, and are termed pluripotent stem cells. Stem cells in the tissues and organs of fetus, infant, and adult have highly reduced potency and committed to produce only progenitor cells for particular tissues. These tissue-specific stem cells are called multipotent stem cells. These tissue-specific/committed multipotent stem cells, when placed in altered environment other than their original niche, can yield cells characteristic of the altered environment. These findings are certainly of potential interest from the clinical, therapeutic perspective. The controversial terminology 'somatic stem cell plasticity' coined by the stem cell community seems to have been proved true. Followings are some of the recent knowledges related to the stem cell. Just as the tissues of our body have their own multipotent stem cells, cancerous tumor has undifferentiated cells known as cancer stem cell (CSC). Each time CSC cleaves, it makes two daughter cells with different fate. One is endowed with immortality, the remarkable ability to divide indefinitely, while the other progeny cell divides occasionally but lives forever. In the cancer tumor, CSC is minority being as few as 3-5% of the tumor mass but it is the culprit behind the tumor-malignancy, metastasis, and recurrence of cancer. CSC is like a master print. As long as the original exists, copies can be made and the disease can persist. If the CSC is destroyed, cancer tumor can't grow. In the decades-long cancer therapy, efforts were focused on the reducing of the bulk of cancerous growth. How cancer therapy is changing to destroy the origin of tumor, the CSC. The next generation of treatments should be to recognize and target the root cause of cancerous growth, the CSC, rather than the reducing of the bulk of tumor, Now the strategy is to find a way to identify and isolate the stem cells. The surfaces of normal as well as the cancer stem cells are studded with proteins. In leukaemia stem cell, for example, protein CD 34 is identified. In the new treatment of cancer disease it is needed to look for protein unique to the CSC. Blocking the stem cell's source of nutrients might be another effective strategy. The mystery of sternness of stem cells has begun to be deciphered. ESC can replicate indefinitely and yet retains the potential to turn into any kind of differentiated cells. Polycomb group protein such as Suz 12 repress most of the regulatory genes which, activated, are turned to be developmental genes. These protein molecules keep the ESC in an undifferentiated state. Many of the regulator genes silenced by polycomb proteins are also occupied by such ESC transcription factors as Oct 4, Sox 2, and Nanog. Both polycomb and transcription factor proteins seem to cooperate to keep the ESC in an undifferentiated state, pluripotent, and self-renewable. A normal prion protein (PrP) is found throughout the body from blood to the brain. Prion diseases such as mad cow disease (bovine spongiform encephalopathy) are caused when a normal prion protein misfolds to give rise to PrP$^{SC}$ and assault brain tissue. Why has human body kept such a deadly and enigmatic protein? Although our body has preserved the prion protein, prion diseases are of rare occurrence. Deadly prion diseases have been intensively studied, but normal prion problems are not. Very few facts on the benefit of prion proteins have been known so far. It was found that PrP was hugely expressed on the stem cell surface of bone marrow and on the cells of neural progenitor, PrP seems to have some function in cell maturation and facilitate the division of stem cells and their self-renewal. PrP also might help guide the decision of neural progenitor cell to become a neuron.

Siberian Sturgeon Oocyte Extract Induces Epigenetic Modifications of Porcine Somatic Cells and Improves Developmental Competence of SCNT Embryos

  • Kim, So-Young;Kim, Tae-Suk;Park, Sang-Hoon;Lee, Mi-Ran;Eun, Hye-Ju;Baek, Sang-Ki;Ko, Yeoung-Gyu;Kim, Sung-Woo;Seong, Hwan-Hoo;Campbell, Keith H.S.;Lee, Joon-Hee
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제27권2호
    • /
    • pp.266-277
    • /
    • 2014
  • Somatic cell nuclear transfer (SCNT) has generally demonstrated that a differentiated cell can convert into a undifferentiated or pluripotent state. In the SCNT experiment, nuclear reprogramming is induced by exposure of introduced donor nuclei to the recipient cytoplasm of matured oocytes. However, because the efficiency of SCNT still remains low, a combination of SCNT technique with the ex-ovo method may improve the normal development of SCNT embryos. Here we hypothesized that treatment of somatic cells with extracts prepared from the germinal vesicle (GV) stage Siberian sturgeon oocytes prior to their use as nuclear donor for SCNT would improve in vitro development. A reversible permeability protocol with $4{\mu}g/mL$ of digitonin for 2 min at $4^{\circ}C$ in order to deliver Siberian sturgeon oocyte extract (SOE) to porcine fetal fibroblasts (PFFs) was carried out. As results, the intensity of H3K9ac staining in PFFs following treatment of SOE for 7 h at $18^{\circ}C$ was significantly increased but the intensity of H3K9me3 staining in PFFs was significantly decreased as compared with the control (p<0.05). Additionally, the level of histone acetylation in SCNT embryos at the zygote stage was significantly increased when reconstructed using SOE-treated cells (p<0.05), similar to that of IVF embryos at the zygote stage. The number of apoptotic cells was significantly decreased and pluripotency markers (Nanog, Oct4 and Sox2) were highly expressed in the blastocyst stage of SCNT embryos reconstructed using SOE-treated cells as nuclear donor (p<0.05). And there was observed a better development to the blastocyst stage in the SOE-treated group (p<0.05). Our results suggested that pre-treatment of cells with SOE could improve epigenetic reprogramming and the quality of porcine SCNT embryos.

MMP2 Gene-735 C/T and MMP9 gene -1562 C/T Polymorphisms in JAK2V617F Positive Myeloproliferative Disorders

  • Sag, Sebnem Ozemri;Gorukmez, Ozlem;Ture, Mehmet;Gorukmez, Orhan;Topak, Ali;Sahinturk, Serdar;Ocakoglu, Gokhan;Gulten, Tuna;Ali, Ridvan;Yakut, Tahsin
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제16권2호
    • /
    • pp.443-449
    • /
    • 2015
  • Background: Myeloproliferative disorders (MPDs) are clonal hematologic malignancies originating at the level of the pluripotent hematopoietic stem cell. Matrix metalloproteases (MMPs) are proteolytic enzymes that contribute to all stages of malignancy progression. Genetic variants in the MMP genes may influence the biological function of these enzymes and change their role in carcinogenesis and progression. To our knowledge, this is the first investigation of associations between the -735 C/T and -1562 C/T polymorphisms in the MMP2 and MMP9 genes, respectively, and the risk of essential thrombocytosis (ET), and polycythemia vera (PV). Materials and Methods: The case-control study included JAK2V617F mutation positive 102 ET and PV patients and 111 controls. Polymorphisms were determined by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and electrophoresis. Results: No statistically significant differences were detected between patient (ET+PV) and control groups regarding genotype distribution for MMP2 gene-735 C/T and MMP9 gene -1562 C/T polymorphisms and C/T allele frequency (p>0.050). Statistically borderline significance was observed between PV and control groups regarding genotype distribution for the MMP9 gene -1562 C/T polymorphism (p=0.050, OR=2.26, 95%Cl=0.99-5.16). Conclusions: Consequently this study supported that CC genotype of MMP9 gene -1562 C/T polymorphism may be related with PV even if with borderline significance.

흉부 질환 진단에서 소홀하기 쉬운 단순 흉부 X-선 사진의 중요성: 종격동 종양을 중심으로 (The Importance of the Simple Chest X-ray Tend to Make Careless Interpretation: with a Case of Mediastinal Tumor)

  • 김은영;임주은;박병훈;윤진영;정지예;손지영;이경종;윤여운;강영애;문진욱;박무석;김영삼;장준;신성관;김세규
    • Tuberculosis and Respiratory Diseases
    • /
    • 제66권6호
    • /
    • pp.482-485
    • /
    • 2009
  • 종격동은 다양한 기관들이 밀집되어 있는 구조라서 종양 등 질병이 발생하더라도 위치상 심장이나 대혈관의 음영과 중첩되어 보이기 때문에 단순 흉부 X-선 촬영상 정상 소견으로 오인할 수 있다. 종격동 종양의 경우 주위조직의 침범 여부나 주위 림프절 크기 이외에도 환자의 연령, 종양의 크기 및 위치, 환자의 증세 유무가 중요한 고려사항이다. 저자들은 흉부 불쾌감이 지속되어 내원한 56세 여자 환자에게서 단순 흉부 X-선 사진의 면밀한 관찰을 통해 기관 분지부의 종격동 종양을 발견한 증례를 통해, 다시 한번 단순 흉부 X-선 촬영의 진단적인 중요성을 임상의들에게 환기시키고자 한다.

인간 배아 줄기세포의 초자화 동결 및 초급속 융해에 관한 연구 (The Study on Vitrification and Ultrarapid Thawing of Human Embryonic Stem Cells)

  • 문신용;박용빈;김희선;성기청;오선경;천대우;서창석;최영민;김정구;이진용;김석현
    • Clinical and Experimental Reproductive Medicine
    • /
    • 제29권1호
    • /
    • pp.13-20
    • /
    • 2002
  • Objective: This study was carried out to establish the effectiveness of the vitrification method and the optimal cryoprotectants in the cryopreservation of human embryonic stem cells (ESC). Materials and Methods: Human ESC clumps established at Seoul National University Hospital (SNUhES 1) were cryopreserved with the vitrification method using the EM grid. EDS and EFS40 were used as vitrification solutions. Results: Between the EDS and EFS40 groups, there was no significant difference in the recovery rate after cryopreservation of human ESC. The formation rates of ESC colonies in the vitrified groups were significantly lower than those in the control ESC group (p<0.05, p<0.05). In addition, the formation rate of ESC colonies in the EDS group was significantly higher than that in the EFS40 group (p<0.05). The ESC colonies in the vitrified groups were significantly smaller after culture duration of 2 and 4 days, respectively, compared with the control ESC group (p<0.1, p<0.05). However, these effects could be reduced to nonsignificant level by the additional culture of ESC colonies. The vitrified human ESC retained the properties of pluripotent cells, including the expression of cell surface. markers for the undifferentiated cells such as alkaline phosphatase and SSEA-4 (stage-specific embryonic antigen-4), and the expression of transcription factor Oct-4 (octamer-binding transcription factor-4), and the normal karyotype. Conclusion: The vitrification method using the EM grid and EDS solution was confirmed to be very effective for the cryopreservation of human ESC.

Effects of Trichostatin A on In vitro Development of Porcine Embryos Derived from Somatic Cell Nuclear Transfer

  • Jeong, Yeon Ik;Park, Chi Hun;Kim, Huen Suk;Jeong, Yeon Woo;Lee, Jong Yun;Park, Sun Woo;Lee, Se Yeong;Hyun, Sang Hwan;Kim, Yeun Wook;Shin, Taeyoung;Hwang, Woo Suk
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제26권12호
    • /
    • pp.1680-1688
    • /
    • 2013
  • Many different approaches have been developed to improve the efficiency of animal cloning by somatic cell nuclear transfer (SCNT), one of which is to modify histone acetylation levels using histone deacetylase inhibitors (HDACi) such as trichostatin A (TSA). In the present study, we examined the effect of TSA on in vitro development of porcine embryos derived from SCNT. We found that TSA treatment (50 nM) for 24 h following oocyte activation improved blastocyst formation rates (to 22.0%) compared with 8.9% in the non-treatment group and total cell number of the blastocysts for determining embryo quality also increased significantly ($88.9{\rightarrow}114.4$). Changes in histone acetylation levels as a result of TSA treatment were examined using indirect immunofluorescence and confocal microscopy scanning. Results showed that the histone acetylation level in TSA-treated embryos was higher than that in controls at both acetylated histone H3 lysine 9 (AcH3K9) and acetylated histone H4 lysine 12 (AcH4K12). Next, we compared the expression patterns of seven genes (OCT4, ID1; the pluripotent genes, H19, NNAT, PEG1; the imprinting genes, cytokeratin 8 and 18; the trophoblast marker genes). The SCNT blastocysts both with and without TSA treatment showed lower levels of OCT4, ID1, cytokeratin 8 and 18 than those of the in vivo blastocysts. In the case of the imprinting genes H19 and NNAT, except PEG1, the SCNT blastocysts both with and without TSA treatment showed higher levels than those of the in vivo blastocysts. Although the gene expression patterns between cloned blastocysts and their in vivo counterparts were different regardless of TSA treatment, it appears that several genes in NT blastocysts after TSA treatment showed a slight tendency toward expression patterns of in vivo blastocysts. Our results suggest that TSA treatment may improve preimplantation porcine embryo development following SCNT.

Transdifferentiation of α-1,3-galactosyltransferase knockout pig bone marrow derived mesenchymal stem cells into pancreatic β-like cells by microenvironment modulation

  • Ullah, Imran;Lee, Ran;Oh, Keon Bong;Hwang, Seongsoo;Kim, Youngim;Hur, Tai-Young;Ock, Sun A
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제33권11호
    • /
    • pp.1837-1847
    • /
    • 2020
  • Objective: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. Methods: The BM-MSCs have been differentiated into pancreatic β-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. Results: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic β-like cells in the N2B27-based media than in the ADMEM-based media. Conclusion: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic β-like cells.

우 수정란의 배양세포들로부터 송아지 생산을 위한 체계 (Systems for Production of Calves from Cultured Bovine Embryonic Cells)

  • 박세필;;정길생;이훈택;윤산현;임진호
    • 한국가축번식학회지
    • /
    • 제18권4호
    • /
    • pp.299-307
    • /
    • 1995
  • 전능성을 지닌 우 수정란 세포배양기술 체계 확립은 가축육종에 중요한 의의를 지닌다. 이러한 체계는 1) 핵치환에 의한 다수의 클론동물 생산에 대한 기전, 2) 형질전환세포를 선발하기 위한 marker의 사용으로 효과적인 유전자 전이체계와 3) site specific 유전자 전이에 대한 기전 또는 homologous DNA서열 재조합에 의한 결손 등에 대한 기전을 이해하는데 이용될 수 있다. 우 수정란세포의 배양은 배반포 내부 세포괴, 상실배와 16∼20세포기로부터 확립하였다. 이들 모든 세포들은 생쥐 배아간 세포형태와 유사하였으며 배양시 분화와 증식에서 다능성을 나타내었다. 배양체계는 미세소적이나 배양용기, 우 도는 설치류 섬유아 세포주를 단기간 배양 또는 장기간 배양방법을 이용하였다. 유사분열시 요구되는 배양체계나 배양액 그리고 분화 억제에 대한 괄목할만한 장점은 아직 밝혀지지 않고 있다. 현재 16∼20 세포기의 배양세포의 전능성에 대해서는 알려져 있지 않다. 배양된 ICM세포 전능성은 27일간 배양한 ICM 세포로부터 4마리의 산자 생산에 의해 입증되었다.

  • PDF

Calcium Ionophore를 이용한 소 배반포로 부터의 배아주 유사세포의 효과적인 분리 (Efficient Establishment of Presumptive Embryonic Stem Cells from Bovine Blastocysts by Exposure to Calcium Ionophore)

  • 김선욱;류재웅;이철상;한용만;박정선;유대열;이경광
    • 한국발생생물학회지:발생과생식
    • /
    • 제3권1호
    • /
    • pp.45-51
    • /
    • 1999
  • 소 배반포로부터 배아주 (embryonic stem, ES) 유사세포를 분리하기 위해서는 영양외배엽 (trophectoderm, TE) 세포를 제거하는 것이 효과적이다. 따라서 본 실험은 효과적으로 TE를 제거하기 위한 calcium ionophore A23187 (CIPA) 처리조건을 확립하고, 분리해낸 ES 유사세포의 in vitro 다능성 (pluripotency)을 검증하고자 수행하였다. CIPA 농도 및 처리시간을 달리 하였을 때 50 $\mu$M에서 25분간 처리한 군이 colony 형성율 (51%)및 10 passage 까지의 배양성적 (4.8%)에서 가장 좋은 결과를 나타내었다. 또한 CIPA를 처리하지 않은 군과의 비교에서도 약 5배의 높은 결과를 보임으로서 본 실험에서 확립된 CIPA 처리조건은 가시적인 toxicity 없이 ES 유사세포의 확립에 이용될 수 있음을 시사하였다. 확립된 ES 유사세포는 heterogeneous한 alkaline phosphatase (AP) 활성을 보여 소 ES 유사세포에 대한 타 보고들과 유사한 결과를 보였다. In vitro 부양배양 (suspension culture)에서는 embryoid body로 분화가 가능하였으며, 약 70% 정도의 euploidism을 보였다. 따라서 본 실험에서 확립된 CIPA의 처리조건이 소 배반포로부터 ES 유사세포를 확립하는데 효과적으로 이용될 수 있음을 확인할 수 있었다.

  • PDF

Effects of Exogenous Insulin-like Growth Factor 2 on Neural Differentiation of Parthenogenetic Murine Embryonic Stem Cells

  • Choi, Young-Ju;Park, Sang-Kyu;Kang, Ho-In;Roh, Sang-Ho
    • Reproductive and Developmental Biology
    • /
    • 제36권1호
    • /
    • pp.33-37
    • /
    • 2012
  • Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.