{"id":{"repo_id":"ajou","oai_identifier":"oai:repository.ajou.ac.kr:201003/8611"},"canonical_url":"https://search.dev.ndltd.org/etd/ajou/oai:repository.ajou.ac.kr:201003/8611","repository":{"repo_id":"ajou","name":"Ajou University","base_url":"http://repository.ajou.ac.kr/oai/request"},"display":{"title":"Role of Wnt signaling in proliferation of human bone marrow-derived mesenchymal stem cells","abstract":"Human mesenchymal stem cell (hMSC) is an attractive source of adult stem cells for the therapeutic use in regenerative medicine. These cells are usually culture expanded prior to their application. However, their proliferation and differentiation potential were gradually decreased during in vitro expansion. Wingless-related MMTV integration site (Wnt) is well known signaling molecules that implicated in mesenchymal differentiation during development, but the role in proliferation of hMSCs is not well known. In this study, we investigated the effect of Wnt signaling molecules on the proliferation of hMSCs. The relative expression of secreted frizzled-related protein 1, an inhibitor for canonical Wnt signaling, was declined with passage, but nuclear β-catenin level was not related to sFRP1. Interestingly, specific knockdown of sFRP1 by shRNA in hMSCs decreased their proliferation rate, whereas recombinant sFRP1 protein accelerates the growth of cells. These findings suggest that sFRP1 has a pivotal role in proliferation of hMSCs during in vitro culture expansion. Additionally, we found that the expression levels of Wnt5a and retinoic-acid orphan related receptor α were gradually increased with passage. These observations suggest that non-canonical Wnt pathway is a potent regulator of hMSC proliferation.","abstract_html":"Human mesenchymal stem cell (hMSC) is an attractive source of adult stem cells for the therapeutic use in regenerative medicine. These cells are usually culture expanded prior to their application. However, their proliferation and differentiation potential were gradually decreased during in vitro expansion. Wingless-related MMTV integration site (Wnt) is well known signaling molecules that implicated in mesenchymal differentiation during development, but the role in proliferation of hMSCs is not well known. In this study, we investigated the effect of Wnt signaling molecules on the proliferation of hMSCs. The relative expression of secreted frizzled-related protein 1, an inhibitor for canonical Wnt signaling, was declined with passage, but nuclear β-catenin level was not related to sFRP1. Interestingly, specific knockdown of sFRP1 by shRNA in hMSCs decreased their proliferation rate, whereas recombinant sFRP1 protein accelerates the growth of cells. These findings suggest that sFRP1 has a pivotal role in proliferation of hMSCs during in vitro culture expansion. Additionally, we found that the expression levels of Wnt5a and retinoic-acid orphan related receptor α were gradually increased with passage. These observations suggest that non-canonical Wnt pathway is a potent regulator of hMSC proliferation.","abstract_has_math":false,"creators":["이, 지은"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["대학원 의생명과학과","201124164"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2013,"date_issued":"2013-12-16","date_published":"2013-12-16","updated_at":"2026-07-24T00:51:42Z","subjects":["MSC","Wnt","sFRP1","Wnt5a"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000015085","000000015085"],"render_values":[{"text":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000015085","href":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000015085","code":true},{"text":"000000015085","href":null,"code":true}]}]},"links":{"outbound_url":"http://repository.ajou.ac.kr/handle/201003/8611","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["대학원 의생명과학과","201124164","이, 지은"]},{"key":"dc:creator","label":"Author","values":["이, 지은"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2013-12-16","2013"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","Theses"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["MSC","Wnt","sFRP1","Wnt5a"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://repository.ajou.ac.kr/handle/201003/8611","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000015085","000000015085"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Human mesenchymal stem cell (hMSC) is an attractive source of adult stem cells for the therapeutic use in regenerative medicine. These cells are usually culture expanded prior to their application. However, their proliferation and differentiation potential were gradually decreased during in vitro expansion. Wingless-related MMTV integration site (Wnt) is well known signaling molecules that implicated in mesenchymal differentiation during development, but the role in proliferation of hMSCs is not well known. In this study, we investigated the effect of Wnt signaling molecules on the proliferation of hMSCs. The relative expression of secreted frizzled-related protein 1, an inhibitor for canonical Wnt signaling, was declined with passage, but nuclear β-catenin level was not related to sFRP1. Interestingly, specific knockdown of sFRP1 by shRNA in hMSCs decreased their proliferation rate, whereas recombinant sFRP1 protein accelerates the growth of cells. These findings suggest that sFRP1 has a pivotal role in proliferation of hMSCs during in vitro culture expansion. Additionally, we found that the expression levels of Wnt5a and retinoic-acid orphan related receptor α were gradually increased with passage. These observations suggest that non-canonical Wnt pathway is a potent regulator of hMSC proliferation.","인간 중간엽 줄기세포는 재생의학 세포치료에 아주 유용한 성체 줄기세포이다. 다양한 질병의 세포치료를 위해 대부분의 연구에서는 이런 중간엽 줄기세포를 배양 한 후에 사용한다. 하지만 생체 외 배양을 하다 보면, 중간엽 줄기세포의 분화능력과 자가 재생능력이 점점 잃어간다. 하여 우리는 줄기세포에서 분화능력은 물론, 자가 재생능력에 큰 영향력이 있는 유전자와 중간엽 줄기세포와의 연관성을 알아보는 연구를 하였다. 우선, 우리는 중간엽 줄기세포를 장기 배양 시 증식능력과 분화능력을 확인한 결과, 시간이 지날수록 분화능력, 증식능력 모두 감소하는 것을 확인하였다. 그 후, 중간엽 줄기세포의 증식능력과 분화능력에 모두 영향이 있다 밝혀진 Wnt 신호전달의 유전자가 중간엽 줄기세포를 장기 배양 시 변화를 확인한 결과, 특이하게도 Wnt의 억제제로 알려져 있는 sFRP1의 감소를 확인하였지만 이 변화는 핵 안의 β-catenin과는 연관이 없었다. 먼저 sFRP1의 기능을 알아보기 위해 세포를 배양할 때 sFRP1 재조합 단백질을 처리하여 세포의 증식의 변화를 살펴본 결과, 아주 미세하게 이지만 증식이 증가한 것을 확인하였다. 반대로 세포에서의 sFRP1의 발현을 억제한 후 세포 증식을 확인한 결과, 발현 억제된 세포의 증식이 현저히 감소한 것을 확인하였다. 이러한 실험으로 sFRP1이 중간엽 줄기세포의 증식 시에 필요한 물질이라는 것을 증명할 수 있었다. 또한 Wnt family에 속하는 Wnt5a의 증가하였고, 그 하위 유전자인 RORα도 역시 증가하였다. 이 실험결과로 Wnt5a 신호 전달이 인간 중간엽 줄기세포 증식의 조절에 역할이 있다는 것을 알게 되었다.","ABSTRACT Ⅰ TABLE OF CONTENTS Ⅱ LIST OF FIGURES Ⅳ LIST OF TABLES Ⅴ LIST OF ABBREVIATION Ⅵ INTRODUCTION 1 MATERIALS AND METHODS 11 Culture of human MSC 11 Growth Kinetics of human MSC 12 In vitro differentiation 12 SA-β-gal staining 12 Reverse transcriptase PCR, quantitative reverse transcriptase PCR 13 Western blot analysis 14 shRNA-mediated Gene Knockdown 14 Recombinant protein treatment methods of human MSCs 16 Immunocytochemistry 16 RESULTS -20 Characterization of hMSC 20 Reverse Transcriptase-PCR and quantitative reverse transcriptase PCR analysis for Wnt signaling molecules 21 Role of sFRP1 in proliferation of h MSC 21 Analysis of Wnt signaling pathway in hMSC 22 DISCUSSION 35 REFERNCES 39 국문요약 46","Master"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Role of Wnt signaling in proliferation of human bone marrow-derived mesenchymal stem cells","Wnt 신호전달의 인간골수유래 중간엽줄기세포 증식에 있어서의 역할"]}]}],"canonical_facts":{"dc:contributor":["대학원 의생명과학과","201124164","이, 지은"],"dc:creator":["이, 지은"],"dc:date":["2013-12-16","2013"],"dc:description":["Human mesenchymal stem cell (hMSC) is an attractive source of adult stem cells for the therapeutic use in regenerative medicine. These cells are usually culture expanded prior to their application. However, their proliferation and differentiation potential were gradually decreased during in vitro expansion. Wingless-related MMTV integration site (Wnt) is well known signaling molecules that implicated in mesenchymal differentiation during development, but the role in proliferation of hMSCs is not well known. In this study, we investigated the effect of Wnt signaling molecules on the proliferation of hMSCs. The relative expression of secreted frizzled-related protein 1, an inhibitor for canonical Wnt signaling, was declined with passage, but nuclear β-catenin level was not related to sFRP1. Interestingly, specific knockdown of sFRP1 by shRNA in hMSCs decreased their proliferation rate, whereas recombinant sFRP1 protein accelerates the growth of cells. These findings suggest that sFRP1 has a pivotal role in proliferation of hMSCs during in vitro culture expansion. Additionally, we found that the expression levels of Wnt5a and retinoic-acid orphan related receptor α were gradually increased with passage. These observations suggest that non-canonical Wnt pathway is a potent regulator of hMSC proliferation.","인간 중간엽 줄기세포는 재생의학 세포치료에 아주 유용한 성체 줄기세포이다. 다양한 질병의 세포치료를 위해 대부분의 연구에서는 이런 중간엽 줄기세포를 배양 한 후에 사용한다. 하지만 생체 외 배양을 하다 보면, 중간엽 줄기세포의 분화능력과 자가 재생능력이 점점 잃어간다. 하여 우리는 줄기세포에서 분화능력은 물론, 자가 재생능력에 큰 영향력이 있는 유전자와 중간엽 줄기세포와의 연관성을 알아보는 연구를 하였다. 우선, 우리는 중간엽 줄기세포를 장기 배양 시 증식능력과 분화능력을 확인한 결과, 시간이 지날수록 분화능력, 증식능력 모두 감소하는 것을 확인하였다. 그 후, 중간엽 줄기세포의 증식능력과 분화능력에 모두 영향이 있다 밝혀진 Wnt 신호전달의 유전자가 중간엽 줄기세포를 장기 배양 시 변화를 확인한 결과, 특이하게도 Wnt의 억제제로 알려져 있는 sFRP1의 감소를 확인하였지만 이 변화는 핵 안의 β-catenin과는 연관이 없었다. 먼저 sFRP1의 기능을 알아보기 위해 세포를 배양할 때 sFRP1 재조합 단백질을 처리하여 세포의 증식의 변화를 살펴본 결과, 아주 미세하게 이지만 증식이 증가한 것을 확인하였다. 반대로 세포에서의 sFRP1의 발현을 억제한 후 세포 증식을 확인한 결과, 발현 억제된 세포의 증식이 현저히 감소한 것을 확인하였다. 이러한 실험으로 sFRP1이 중간엽 줄기세포의 증식 시에 필요한 물질이라는 것을 증명할 수 있었다. 또한 Wnt family에 속하는 Wnt5a의 증가하였고, 그 하위 유전자인 RORα도 역시 증가하였다. 이 실험결과로 Wnt5a 신호 전달이 인간 중간엽 줄기세포 증식의 조절에 역할이 있다는 것을 알게 되었다.","ABSTRACT Ⅰ TABLE OF CONTENTS Ⅱ LIST OF FIGURES Ⅳ LIST OF TABLES Ⅴ LIST OF ABBREVIATION Ⅵ INTRODUCTION 1 MATERIALS AND METHODS 11 Culture of human MSC 11 Growth Kinetics of human MSC 12 In vitro differentiation 12 SA-β-gal staining 12 Reverse transcriptase PCR, quantitative reverse transcriptase PCR 13 Western blot analysis 14 shRNA-mediated Gene Knockdown 14 Recombinant protein treatment methods of human MSCs 16 Immunocytochemistry 16 RESULTS -20 Characterization of hMSC 20 Reverse Transcriptase-PCR and quantitative reverse transcriptase PCR analysis for Wnt signaling molecules 21 Role of sFRP1 in proliferation of h MSC 21 Analysis of Wnt signaling pathway in hMSC 22 DISCUSSION 35 REFERNCES 39 국문요약 46","Master"],"dc:format":["application/pdf"],"dc:identifier":["http://repository.ajou.ac.kr/handle/201003/8611","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000015085","000000015085"],"dc:language":["en"],"dc:subject":["MSC","Wnt","sFRP1","Wnt5a"],"dc:title":["Role of Wnt signaling in proliferation of human bone marrow-derived mesenchymal stem cells","Wnt 신호전달의 인간골수유래 중간엽줄기세포 증식에 있어서의 역할"],"dc:type":["Thesis","Theses"]},"updated_at":"2026-07-24T00:51:42Z"}