Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Baculovirus transduction of human mesenchymal stem cell-derived progenitor cells: variation of transgene expression with cellular differentiation states

Gene Therapy volume 13, pages 1471–1479 (2006)Cite this article

Abstract

We have previously demonstrated that baculovirus can efficiently transduce human mesenchymal stem cells (MSCs). In this study, we further demonstrated, for the first time, that baculovirus can transduce adipogenic, chondrogenic and osteogenic progenitors originating from MSCs. The transduction efficiency (21–90%), transgene expression level and duration (7–41 days) varied widely with the differentiation lineages and stages of the progenitors, as determined by flow cytometry. The variation stemmed from differential transgene transcription (as revealed by real-time reverse transcription-polymerase chain reaction), rather than from variability in virus entry or cell cycle (as determined by quantitative real-time PCR and flow cytometry). Nonetheless, the baculovirus-transduced cells remained capable of differentiating into adipogenic, osteogenic and chondrogenic pathways. The susceptibility to baculovirus transduction was higher for adipogenic and osteogenic progenitors, but was lower for chondrogenic progenitors. In particular, the duration of transgene expression was prolonged in the transduced adipogenic and osteogenic progenitors (as opposed to the MSCs), implicating the possibility of extending transgene expression via a proper transduction strategy design. Taken together, baculovirus may be an attractive alternative to genetically modify adipogenic and osteogenic progenitors in the ex vivo setting for cell therapy or tissue engineering.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Caplan AI, Bruder SP . Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. Trends Mol Med 2001; 7: 259–264.

    Article  CAS  PubMed  Google Scholar 

  2. Bruder SP, Jaiswal N, Haynesworth SE . Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. J Cell Biochem 1997; 64: 278–294.

    Article  CAS  PubMed  Google Scholar 

  3. Chan J, O'Donoghue K, de la Fuente J, Roberts IA, Kumar S, Morgan JE et al. Human fetal mesenchymal stem cells as vehicles for gene delivery. Stem Cells 2005; 23: 93–102.

    Article  CAS  PubMed  Google Scholar 

  4. Marx JC, Allay JA, Persons DA, Nooner SA, Hargrove PW, Kelly PF et al. High-efficiency transduction and long-term gene expression with a murine stem cell retroviral vector encoding the green fluorescent protein in human marrow stromal cells. Hum Gene Ther 1999; 10: 1163–1173.

    Article  CAS  PubMed  Google Scholar 

  5. Tsuda H, Wada T, Ito Y, Uchida H, Dehari H, Nakamura K et al. Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector. Mol Ther 2003; 7: 354–365.

    Article  CAS  PubMed  Google Scholar 

  6. Gazit D, Turgeman G, Kelley P, Wang E, Jalenak M, Zilberman Y et al. Engineered pluripotent mesenchymal cells integrate and differentiate in regenerating bone: a novel cell-mediated gene therapy. J Gene Med 1999; 1: 121–133.

    Article  CAS  PubMed  Google Scholar 

  7. Lee K, Majumdar MK, Buyaner D, Hendricks JK, Pittenger MF, Mosca JD . Human mesenchymal stem cells maintain transgene expression during expansion and differentiation. Mol Ther 2001; 3: 857–866.

    Article  CAS  PubMed  Google Scholar 

  8. Asahara T, Kalka C, Isner JM . Stem cell therapy and gene transfer for regeneration. Gene Therapy 2000; 7: 451–457.

    Article  CAS  PubMed  Google Scholar 

  9. Verma IM, Somia N . Gene therapy–promises, problems and prospects. Nature 1997; 389: 239–242.

    Article  CAS  PubMed  Google Scholar 

  10. Kost TA, Condreay JP . Recombinant baculoviruses as mammalian cell gene delivery vectors. Trends Biotechnol 2002; 20: 173–180.

    Article  CAS  PubMed  Google Scholar 

  11. Hu Y-C . Baculovirus as a highly efficient expression vector in insect and mammalian cells. Acta Pharmacol Sin 2005; 26: 405–416.

    Article  CAS  PubMed  Google Scholar 

  12. Kost TA, Condreay JP, Jarvis DL . Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 2005; 23: 567–575.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ho Y-C, Chen H-C, Wang K-C, Hu Y-C . Highly efficient baculovirus-mediated gene transfer into rat chondrocytes. Biotechnol Bioeng 2004; 88: 643–651.

    Article  CAS  PubMed  Google Scholar 

  14. Wagle M, Jesuthasan S . Baculovirus-mediated gene expression in zebrafish. Mar Biotechnol 2003; 5: 58–63.

    Article  CAS  Google Scholar 

  15. Airenne KJ, Hiltunen MO, Turunen MP, Turunen AM, Laitinen OH, Kulomaa MS et al. Baculovirus-mediated periadventitial gene transfer to rabbit carotid artery. Gene Therapy 2000; 7: 1499–1504.

    Article  CAS  PubMed  Google Scholar 

  16. Tani H, Limn CK, Yap CC, Onishi M, Nozaki M, Nishimune Y et al. In vitro and in vivo gene delivery by recombinant baculoviruses. J Virol 2003; 77: 9799–9808.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ho Y-C, Chung Y-C, Hwang S-M, Wang K-C, Hu Y-C . Transgene expression and differentiation of baculovirus-transduced human mesenchymal stem cells. J Gene Med 2005; 7: 860–868.

    Article  CAS  PubMed  Google Scholar 

  18. Sutton RE, Reitsma MJ, Uchida N, Brown PO . Transduction of human progenitor hematopoietic stem cells by human immunodeficiency virus type 1-based vectors is cell cycle dependent. J Virol 1999; 73: 3649–3660.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Seidman MA, Hogan SM, Wendland RL, Worgall S, Crystal RG, Leopold PL . Variation in adenovirus receptor expression and adenovirus vector-mediated transgene expression at defined stages of the cell cycle. Mol Ther 2001; 4: 13–21.

    Article  CAS  PubMed  Google Scholar 

  20. Ponnazhagan S, Mukherjee P, Wang XS, Qing K, Kube DM, Mah C et al. Adeno-associated virus type 2-mediated transduction in primary human bone marrow-derived CD34+ hematopoietic progenitor cells: donor variation and correlation of transgene expression with cellular differentiation. J Virol 1997; 71: 8262–8267.

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Blum JS, Parrott MB, Mikos AG, Barry MA . Early osteoblastic differentiation induced by dexamethasone enhances adenoviral gene delivery to marrow stromal cells. J Orthop Res 2004; 22: 411–416.

    Article  CAS  PubMed  Google Scholar 

  22. Mizuguchi H, Sasaki T, Kawabata K, Sakurai F, Hayakawa T . Fiber-modified adenovirus vectors mediate efficient gene transfer into undifferentiated and adipogenic-differentiated human mesenchymal stem cells. Biochem Biophys Res Commun 2005; 332: 1101–1106.

    Article  CAS  PubMed  Google Scholar 

  23. Hung SC, Lu CY, Shyue SK, Liu HC, Ho LL . Lineage differentiation-associated loss of adenoviral susceptibility and coxsackie-adenovirus receptor expression in human mesenchymal stem cells. Stem Cells 2004; 22: 1321–1329.

    Article  PubMed  Google Scholar 

  24. Duisit G, Saleun S, Douthe S, Barsoum J, Chadeuf G, Moullier P . Baculovirus vector requires electrostatic interactions including heparan sulfate for efficient gene transfer in mammalian cells. J Gene Med 1999; 1: 93–102.

    Article  CAS  PubMed  Google Scholar 

  25. Tani H, Nishijima M, Ushijima H, Miyamura T, Matsuura Y . Characterization of cell-surface determinants important for baculovirus infection. Virology 2001; 279: 343–353.

    Article  CAS  PubMed  Google Scholar 

  26. Sandig V, Hofmann C, Steinert S, Jennings G, Schlag P, Strauss M . Gene transfer into hepatocytes and human liver tissue by baculovirus vectors. Hum Gene Ther 1996; 7: 1937–1945.

    Article  CAS  PubMed  Google Scholar 

  27. van Loo ND, Fortunati E, Ehlert E, Rabelink M, Grosveld F, Scholte BJ . Baculovirus infection of nondividing mammalian cells: mechanisms of entry and nuclear transport of capsids. J Virol 2001; 75: 961–970.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Salminen M, Airenne KJ, Rinnankoski R, Reimari J, Valilehto O, Rinne J et al. Improvement in nuclear entry and transgene expression of baculoviruses by disintegration of microtubules in human hepatocytes. J Virol 2005; 79: 2720–2728.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Vroemen M, Weidner N, Blesch A . Loss of gene expression in lentivirus- and retrovirus-transduced neural progenitor cells is correlated to migration and differentiation in the adult spinal cord. Exp Neurol 2005; 195: 127–139.

    Article  CAS  PubMed  Google Scholar 

  30. Madry H, Zurakowski D, Trippel SB . Overexpression of human insulin-like growth factor-I promotes new tissue formation in an ex vivo model of articular chondrocyte transplantation. Gene Therapy 2001; 8: 1443–1449.

    Article  CAS  PubMed  Google Scholar 

  31. Obradovic B, Martin I, Padera RF, Treppo S, Freed LE, Vunjak-Novakovic G . Integration of engineered cartilage. J Orthop Res 2001; 19: 1089–1097.

    Article  CAS  PubMed  Google Scholar 

  32. Abe T, Takahashi H, Hamazaki H, Miyano-Kurosaki N, Matsuura Y, Takaku H . Baculovirus induces an innate immune response and confers protection from lethal influenza virus infection in mice. J Immunol 2003; 171: 1133–1139.

    Article  CAS  PubMed  Google Scholar 

  33. Beck NB, Sidhu JS, Omiecinski CJ . Baculovirus vectors repress phenobarbital-mediated gene induction and stimulate cytokine expression in primary cultures of rat hepatocytes. Gene Therapy 2000; 7: 1274–1283.

    Article  CAS  PubMed  Google Scholar 

  34. Erices A, Conget P, Minguell JJ . Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol 2000; 109: 235–242.

    Article  CAS  PubMed  Google Scholar 

  35. Hung SC, Chen NJ, Hsieh SL, Li H, Ma HL, Lo WH . Isolation and characterization of size-sieved stem cells from human bone marrow. Stem Cells 2002; 20: 249–258.

    Article  PubMed  Google Scholar 

  36. Liu P, Kalajzic I, Stover ML, Rowe DW, Lichtler AC . Human bone marrow stromal cells are efficiently transduced by vesicular stomatitis virus-pseudotyped retrovectors without affecting subsequent osteoblastic differentiation. Bone 2001; 29: 331–335.

    Article  CAS  PubMed  Google Scholar 

  37. Aaron RK, Wang S, Ciombor DM . Upregulation of basal TGFβ1 levels by EMF coincident with chondrogenesis – implications for skeletal repair and tissue engineering. J Orthop Res 2002; 20: 233–240.

    Article  CAS  PubMed  Google Scholar 

  38. Hu Y-C, Tsai C-T, Chung Y-C, Lu J-T, Hsu JT-A . Generation of chimeric baculovirus with histidine-tags displayed on the envelope and its purification using immobilized metal affinity chromatography. Enzyme Microb Technol 2003; 33: 445–452.

    Article  CAS  Google Scholar 

  39. Chung Y-C, Liu H-J, Hu Y-C . Facile monitoring of avian reovirus σB expression and purification processes by tagged green fluorescent protein. Enzyme Microb Technol 2004; 35: 494–500.

    Article  CAS  Google Scholar 

  40. Dhar AK, Roux MM, Klimpel KR . Quantitative assay for measuring the Taura syndrome virus and yellow head virus load in shrimp by real-time RT-PCR using SYBR Green chemistry. J Virol Methods 2002; 104: 69–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We gratefully acknowledge the financial support from the National Health Research Institutes (NHRI-EX94-9412EI), Taiwan.

Author information

Authors and Affiliations

  1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan

    Y-C Ho, H-P Lee, W-H Lo, H-C Chen, C-K Chung & Y-C Hu

  2. Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan

    S-M Hwang

Authors
  1. Y-C Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H-P Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S-M Hwang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. W-H Lo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H-C Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C-K Chung
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Y-C Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Y-C Hu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ho, YC., Lee, HP., Hwang, SM. et al. Baculovirus transduction of human mesenchymal stem cell-derived progenitor cells: variation of transgene expression with cellular differentiation states. Gene Ther 13, 1471–1479 (2006). https://doi.org/10.1038/sj.gt.3302796

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302796

Keywords

This article is cited by

Search

Advanced search

Quick links