A Role of Cell Adhesion Molecules and Gelatinases in Human Serum-Induced Aggregation of Human Eyelid-Derived Stem Cells In Vitro
4
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Human serum (HS) has been reported to induce aggregation of human eyelid adipose-derived stem cells (HEACs) during high-density culture in vitro. The present study focused on the role of cell adhesion molecules and gelatinases during HS-induced aggregation of HEACs. HS-induced aggregation occurred between 9-15 days of culture. Cells aggregated by HS medium (HS-agg) showed stronger expression of α2, α2B, αX, and CEACAM1 genes compared to non-aggregated cells in HS medium (HS-ex) or in control FBS-cultured cells. HS-agg were distinctly labeled with antibodies against α2, α2B, and αX proteins. Western blot results demonstrated that the two integrin proteins were greatly expressed in HS-agg compared to HS-ex and control FBS-cultured cells. Treatment of HEACs with anti-integrin α2 antibody during culture in HS medium delayed aggregation formation. HS-agg exhibited strong expression of MMP1 and MMP9 compared to HS-ex or FBS-cultured cells. Conditioned media from HS-culture showed remarkable increase of MMP9 gelatinolytic activity in comparison to those from FBS-culture. However, there was no change of TIMP mRNA expression in relation to the HS-induced aggregation. Based on these results, it is suggested that integrin α2, α2B, and αX, and MMP9 might play an important role in the HS-induced aggregation of HEACs.
Abstract
Human serum (HS) has been reported to induce aggregation of human eyelid adipose-derived stem cells (HEACs) during high-density culture in vitro. The present study focused on the role of cell adhesion molecules and gelatinases during HS-induced aggregation of HEACs. HS-induced aggregation occurred between 9-15 days of culture. Cells aggregated by HS medium (HS-agg) showed stronger expression of α2, α2B, αX, and CEACAM1 genes compared to non-aggregated cells in HS medium (HS-ex) or in control FBS-cultured cells. HS-agg were distinctly labeled with antibodies against α2, α2B, and αX proteins. Western blot results demonstrated that the two integrin proteins were greatly expressed in HS-agg compared to HS-ex and control FBS-cultured cells. Treatment of HEACs with anti-integrin α2 antibody during culture in HS medium delayed aggregation formation. HS-agg exhibited strong expression of MMP1 and MMP9 compared to HS-ex or FBS-cultured cells. Conditioned media from HS-culture showed remarkable increase of MMP9 gelatinolytic activity in comparison to those from FBS-culture. However, there was no change of TIMP mRNA expression in relation to the HS-induced aggregation. Based on these results, it is suggested that integrin α2, α2B, and αX, and MMP9 might play an important role in the HS-induced aggregation of HEACs.
참고문헌 (29)
- Bianco P, "Mesenchymal stem cells: revisiting history, concepts, and assays", Cell Stem Cell, 2, pp.313~319 (2008)
- Bieback K, "Altered gene expression in human adipose stem cells cultured with fetal bovine serum compared to human supplements", Tissue Eng Part A, 16, pp.3467~3484 (2010)
- Bieback K, "Human alternatives to fetal bovine serum for the expansion of mesenchymal stromal cells from bone marrow", Stem Cells, 27, pp.2331~2341 (2009)
- Bieback K, "Replicative aging and differentiation potential of human adipose tissue-derived mesenchymal stromal cells expanded in pooled human or fetal bovine serum", Cytotherapy, 14, pp.570~583 (2012)
- Brooke G, "Molecular trafficking mechanisms of multipotent mesenchymal stem cells derived from human bone marrow and placenta", Stem Cells Dev, 17, pp.929~940 (2008)
- Carrion B, "Bone marrow-derived mesenchymal stem cells enhance angiogenesis via their α6β1 integrin receptor", Exp Cell Res, 319, pp.2964~2476 (2013)
- Cheng NC, "Short-term spheroid formation enhances the regenerative capacity of adipose-derived stem cells by promoting stemness, angiogenesis, and chemotaxis", Stem Cells Transl Med, 2, pp.584~594 (2013)
- De Becker A, "Migration of culture-expanded human mesenchymal stem cells through bone marrow endothelium is regulated by matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3", Haematologica, 92, pp.440~449 (2007)
- Dumin JA, "Pro-collagenase-1 (matrix metalloproteinase-1) binds the alpha(2)beta(1) integrin upon release from keratinocytes migrating on type I collagen", J Biol Chem, 276, pp.29368~29374 (2001)
- Frith JE, "Tailored integrin-extracellular matrix interactions to direct human mesenchymal stem cell differentiation", Stem Cells Dev, 21, pp.2442~2456 (2012)
- Gellersen B, "Human endometrial stromal cell-trophoblast interactions: mutual stimulation of chemotactic migration and promigratory roles of cell surface molecules CD82 and CEACAM1", Biol Reprod, 88, pp.80~ (2013)
- Granero-Moltó F, "Regenerative effects of transplanted mesenchymal stem cells in fracture healing", Stem Cells, 27, pp.1887~1898 (2009)
- Jarvis GE, "A role for adhesion and degranulationpromoting adapter protein in collagen-induced platelet activation mediated via integrin α(2) β(1)", J Thromb Haemost, 10, pp.268~277 (2012)
- Kaneider NC, "Enhancement of fibrinogentriggered pro-coagulant activation of monocytes in vitro by matrix metalloproteinase-9", Thromb J, 8, pp.1~7 (2010)
- Kang HM, "Insulinsecreting cells from human eyelid -derived stem cells alleviate type I diabetes in immunocompetent mice", Stem Cells Dev, 27, pp.1999~2008 (2009)
- Kawai K, "Matrix metalloproteinase-9 contributes to the mobilization of bone marrow cells in the injured liver", Cell Transplant, 21, pp.453~464 (2012)
- Kocaoemer A, "Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue", Stem Cells, 25, pp.1270~1278 (2007)
- McBane RD 2nd, "Platelet recruitment to venous stent thrombi", J Thromb Thrombolysis, 36, pp.442~427 (2013)
- Mizuno H, "Concise review: Adipose-derived stem cells as a novel tool for future regenerative medicine", Stem Cells, 30, pp.804~810 (2012)
- Neumann FJ, "Effect of glycoprotein IIb", IIIa receptor blockade on recovery of coronary flow and left ventricular function after the placement of coronary-artery stents in acute myocardial infarction, 1998, pp.98, 2695~2701
- Prowse AB, "Stem cell integrins: implications for ex-vivo culture and cellular therapies", Stem Cell Res, 6, pp.1~12 (2011)
- Ries C, "MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines", Blood, 109, pp.4055~4063 (2007)
- Sarkar D, "Engineered cell homing", Blood, 118, pp.184~191 (2011)
- Schnickmann S, "AP-1-controlled hepatocyte growth factor activation promotes keratinocyte migration via CEACAM1 and urokinase plasminogen activator", urokinase plasminogen receptor, 2009, pp.129, 1140~1148
- 송연화, "Aggregation of Human Eyelid Adipose-derived Stem Cells by Human Body Fluids", 발생과 생식, 16(4), pp.339~351 (2012), 한국발생생물학회
- Taniguchi Ishikawa E, "Klf5 controls bone marrow homing of stem cells and progenitors through Rab5-mediated β1", β2-integrin trafficking, 2013, pp.4, 1660~
- Tateishi K, "Comparison of human serum with fetal bovine serum for expansion and differentiation of human synovial MSC: potential feasibility for clinical applications", Cell Transplant, 17, pp.549~557 (2008)
- Uccelli A, "Mesenchymal stem cells in health and disease", Nat Rev Immunology, 8, pp.726~736 (2008)
- Zhang H, "Role of CEACAM1 and CEACAM20 in an in vitro model of prostate morphogenesis", PLoS One, 8, pp.e53359~ (2013)