Journal Mobile Options
Table of Contents
Vol. 182, No. 1, 2006
Issue release date: April 2006
Cells Tissues Organs 2006;182:1–11
(DOI:10.1159/000091713)

Characterization of Osteogenically Induced Adipose Tissue-Derived Precursor Cells in 2-Dimensional and 3-Dimensional Environments

Leong D.T. · Khor W.M. · Chew F.T. · Lim T.-C. · Hutmacher D.W.
aDepartment of Biological Sciences, bDepartment of Mechanical Engineering, cDivision of Bioengineering and dDepartment of Orthopedic Surgery, National University of Singapore and eDivision of Plastic Surgery, National University Hospital, Singapore, Singapore

Individual Users: Register with Karger Login Information

Please create your User ID & Password





Contact Information











I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in

Abstract

Earlier reports on a putative precursor cell population in adipose tissue showed differentiation along several mesodermal lineages, leading some to think that adipose tissue can be a source of cells applicable in regenerative medicine. However, characterizations of these adipose-derived precursor cells (ADPC) in the 3-dimensional (3-D) environment, especially within the area of bone-specific composite scaffolds, have been lacking. In this study, ADPC plated on culture flasks or seeded on medical grade polycaprolactone-tricalcium phosphate scaffolds (mPCL-CaP) were able to differentiate along the osteogenic lineages in both 2-D and 3-D environments as assessed with immunohistochemistry of osteo-related proteins, reverse transcriptase-polymerase chain reactions and alkaline phosphatase assay. The mPCL-CaP scaffolds provided adipose-derived cells (ADC) with a suitable environment as determined by DNA and metabolic assays, light, confocal and scanning electron microscopy. Flow cytometry revealed ADC to be CD29+, CD44+, CD73+, CD90+ and CD14–, CD31–, CD34–, CD45–, CD71–, and therefore showed the absence of hematopoietic stem cells but possibly the presence of pericytes and mescenchymal stem cells with osteogenic potential. The results of this study demonstrated the potential of using ADPC in combination with mPCL-CaP scaffolds for bone regenerative medicine.



Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Ahn, S.J., J. Costa, J.R. Emanuel (1996) PicoGreen quantitation of DNA: effective evaluation of samples pre- or post-PCR. Nucleic Acids Res 24: 2623–2625.
  2. Aust, L., B. Devlin, S.J. Foster, Y.D. Halvorsen, K. Hicok, T. du Laney, A. Sen, G.D. Willingmyre, J.M. Gimble (2004) Yield of human adipose-derived adult stem cells from liposuction aspirates. Cytotherapy 6: 7–14.
  3. Awad, H.A., M.Q. Wickham, H.A. Leddy, J.M. Gimble, F. Guilak (2004) Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. Biomaterials 25: 3211–3222.
  4. Collett G, Wood A, Alexander MY, Varnum BC, Boot-Handford RP, Ohanian V, Ohanian J, Fridell YW, Canfield AE (2003) Receptor tyrosine kinase Axl modulates the osteogenic differentiation of pericytes. Circ Res 92: 1123–1129.
  5. Cowan CM, Shi YY, Aalami OO, Chou YF, Mari C, Thomas R, Quarto N, Contag CH, Wu B, Longaker MT (2004) Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol 22: 560–567.
  6. Delany AM, Amling M, Priemel M, Howe C, Baron R, Canalis E (2000) Osteopenia and decreased bone formation in osteonectin-deficient mice. J Clin Invest 105: 1325.
  7. Doherty MJ, Ashton BA, Walsh S, Beresford JN, Grant ME, Canfield AE (1998) Vascular pericytes express osteogenic potential in vitro and in vivo. J Bone Miner Res 13: 828–838.
  8. Dragoo JL, Choi JY, Lieberman JR, Huang J, Zuk PA, Zhang J, Hedrick MH, Benhaim P (2003) Bone induction by BMP-2 transduced stem cells derived from human fat. J Orthop Res 21: 622–629.
  9. Frank O, Heim M, Jakob M, Barbero A, Schafer D, Bendik I, Dick W, Heberer M, Martin I (2002) Real-time quantitative RT-PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro. J Cell Biochem 85: 737–746.
  10. Gaustad KG, Boquest AC, Anderson BE, Gerdes AM, Collas P (2004) Differentiation of human adipose tissue stem cells using extracts of rat cardiomyocytes. Biochem Biophys Res Commun 314: 420–427.
  11. Green D, Walsh D, Mann S, Oreffo RO (2002) The potential of biomimesis in bone tissue engineering: lessons from the design and synthesis of invertebrate skeletons. Bone 30: 810–815.
  12. Gronthos S, Franklin DM, Leddy HA, Robey PG, Storms RW, Gimble JM (2001) Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol 189: 54–63.
  13. Gurevitch O, Kurkalli BG, Prigozhina T, Kasir J, Gaft A, Slavin S (2003) Reconstruction of cartilage, bone, and hematopoietic microenvironment with demineralized bone matrix and bone marrow cells. Stem Cells 21: 588–597.
  14. Hagedorn M, Balke M, Schmidt A, Bloch W, Kurz H, Javerzat S, Rousseau B, Wilting J, Bikfalvi A (2004) VEGF coordinates interaction of pericytes and endothelial cells during vasculogenesis and experimental angiogenesis. Dev Dyn 230: 23–33.
  15. Hicok KC, Du Laney TV, Zhou YS, Halvorsen YD, Hitt DC, Cooper LF, Gimble JM (2004) Human adipose-derived adult stem cells produce osteoid in vivo. Tissue Eng 10: 371–380.
  16. Huss R, Lange C, Weissinger EM, Kolb HJ, Thalmeier K (2000) Evidence of peripheral blood-derived, plastic-adherent CD34(–/low) hematopoietic stem cell clones with mesenchymal stem cell characteristics. Stem Cells 18: 252–260.
  17. Hutmacher DW, Garcia AJ (2005) Scaffold-based bone engineering by using genetically modified cells. Gene 347: 1–10.
  18. Hutmacher DW, Risbud M, Sittinger M (2004) Evolution of computer aided design and advanced manufacturing in scaffold research. Trends Biotechnol 22: 354–362.
  19. Hutmacher, D.W., D. Rohner, P. See (2003) Craniofacial bone tissue engineering using medical imaging, computational modeling, rapid prototyping, bioresorbable scaffolds and bone marrow aspirates; in Reis, R., D. Cohn (eds): Polymer Based Systems in Tissue Engineering, Replacement and Regeneration. Dordrecht, Kluwer Academic Publishers, pp 333–356.
  20. Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM (2002) Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418: 41–49.
  21. Justesen J, Pedersen SB, Stenderup K, Kassem M (2004) Subcutaneous adipocytes can differentiate into bone-forming cells in vitro and in vivo. Tissue Eng 10: 381–391.
  22. Knabe C, Berger G, Gildenhaar R, Meyer J, Howlett CR, Markovic B, Zreiqat H (2004) Effect of rapidly resorbable calcium phosphates and a calcium phosphate bone cement on the expression of bone-related genes and proteins in vitro. J Biomed Mater Res 69: 145–154.
  23. Kuwana M, Okazaki Y, Kodama H, Izumi K, Yasuoka H, Ogawa Y, Kawakami Y, Ikeda Y (2003) Human circulating CD14+ monocytes as a source of progenitors that exhibit mesenchymal cell differentiation. J Leukoc Biol 74: 833–845.
  24. Lee HS, Huang GT, Chiang H, Chiou LL, Chen MH, Hsieh CH, Jiang CC (2003) Multipotential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis. Stem Cells 21: 190–199.
  25. Lee RH, Kim B, Choi I, Kim H, Choi HS, Suh K, Bae YC, Jung JS (2004) Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue. Cell Physiol Biochem 14: 311–324.
  26. Lendeckel S, Jodicke A, Christophis P, Heidinger K, Wolff J, Fraser JK, Hedrick MH, Berthold L, Howaldt HP (2004) Autologous stem cells (adipose) and fibrin glue used to treat widespread traumatic calvarial defects: case report. J Craniomaxillofac Surg 32: 370–373.
  27. Lis GJ, Rokita E, Podolec P, Pfitzner R, Dziatkowiak A, Cichocki T (2003) Mineralization and organic phase modifications as contributory factors of accelerated degeneration in homograft aortic valves. J Heart Valve Dis 12: 741–751.
  28. Mann V, Hobson EE, Li B, Stewart TL, Grant SF, Robins SP, Aspden RM, Ralston SH (2001) A COL1A1 Sp1 binding site polymorphism predisposes to osteoporotic fracture by affecting bone density and quality. J Clin Invest 107: 899–907.
  29. Martina M, Leong TW, Gossens K, et al (2003) The isolation of precursor cell surface markers positive cells in a heterogeneous cell population derived from human adipose tissue. Tissue Engineering Society International, 6th Annual International Conference and Exposition, Orlando.
  30. Mizuno H, Zuk PA, Zhu M, Lorenz HP, Benhaim P, Hedrick MH (2002) Myogenic differentiation by human processed lipoaspirate cells. Plast Reconstr Surg 109: 199–209.
  31. Negrin RS, Atkinson K, Leemhuis T, Hanania E, Juttner C, Tierney K, Hu WW, Johnston LJ, Shizurn JA, Stockerl-Goldstein KE, Blume KG, Weissman IL, Bower S, Baynes R, Dansey R, Karanes C, Peters W, Klein J (2000) Transplantation of highly purified CD34+Thy-1+ hematopoietic stem cells in patients with metastatic breast cancer. Biol Blood Marrow Transplant 6: 262–271.
  32. Neuss S, Becher E, Woltje M, Tietze L, Jahnen-Dechent W (2004) Functional expression of HGF and HGF receptor/c-met in adult human mesenchymal stem cells suggests a role in cell mobilization, tissue repair, and wound healing. Stem Cells 22: 405–414.
  33. Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, Pickel J, McKay R, Nadal-Ginard B, Bodine DM, Leri A, Anversa P (2001) Bone marrow cells regenerate infarcted myocardium. Nature 410: 701–705.
  34. Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwinse S, Lian JB, Stein GS (1990) Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J Cell Physiol 143: 420–430.
  35. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–147.
  36. Rai B, Teoh SH, Hutmacher DW, Cao T, Ho KH (2005) Novel PCL-based honeycomb scaffolds as drug delivery systems for rhBMP-2. Biomaterials 26: 3739–3748.
  37. Rauch F, Glorieux FH (2004) Osteogenesis imperfecta. Lancet 363: 1377–1385.
  38. Reyes M, Lund T, Lenvik T, Aguiar D, Koodie L, Verfaillie CM (2001) Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98: 2615–2625.
  39. Rodan GA, Noda M (1991) Gene expression in osteoblastic cells. Crit Rev Eukaryot Gene Expr 1: 85–98.
  40. Rotter N, Aigner J, Naumann A, Planck H, Hammer C, Burmester G, Sittinger M (1998) Cartilage reconstruction in head and neck surgery: comparison of resorbable polymer scaffolds for tissue engineering of human septal cartilage. J Biomed Mater Res 42: 347–356.
  41. Safford KM, Hicok KC, Safford SD, Halvorsen YD, Wilkison WO, Gimble JM, Rice HE (2002) Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem Biophys Res Commun 294: 371–379.
  42. Sahota O, Mundey MK, San P, Godber IM, Lawson N, Hosking DJ (2004) The relationship between vitamin D and parathyroid hormone: calcium homeostasis, bone turnover, and bone mineral density in postmenopausal women with established osteoporosis. Bone 35: 312–319.
  43. Schantz JT, Teoh SH, Lim TC, Endres M, Lam CX, Hutmacher DW (2003) Repair of calvarial defects with customized tissue-engineered bone grafts. I. Evaluation of osteogenesis in a three-dimensional culture system. Tissue Eng 9(suppl 1): S113–S126.

    External Resources

  44. Schecroun N, Delloye C (2004) In vitro growth and osteoblastic differentiation of human bone marrow stromal cells supported by autologous plasma. Bone 35: 517–524.
  45. Silva WA Jr, Covas DT, Panepucci RA, Proto-Siqueira R, Siufi JL, Zanette DL, Santos AR, Zago MA (2003) The profile of gene expression of human marrow mesenchymal stem cells. Stem Cells 21: 661–669.
  46. Sun S, Guo Z, Xiao X, Liu B, Liu X, Tang PH, Mao N (2003) Isolation of mouse marrow mesenchymal progenitors by a novel and reliable method. Stem Cells 21: 527–535.
  47. Weidt C, Niggemann B, Hatzmann W, Zanker KS, Dittmar T (2004) Differential effects of culture conditions on the migration pattern of stromal cell-derived factor-stimulated hematopoietic stem cells. Stem Cells 22: 890–896.
  48. Weissman IL, Baltimore D (2001) Disappearing stem cells, disappearing science. Science 292: 601.
  49. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13: 4279–4295.
  50. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7: 211–228.


Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50