In vivo bioluminescence imaging for viable human neural stem cells incorporated within in situ gelatin hydrogels

Do Won Hwang, Kyung Min Park, Hye kyung Shim, Yeona Jin, Hyun Jeong Oh, So Won Oh, Song Lee, Hyewon Youn, Yoon Ki Joung, Hong J. Lee, Seung U. Kim, Ki Dong Park, Dong Soo Lee

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Background: Three-dimensional (3D) hydrogel-based stem cell therapies contribute to enhanced therapeutic efficacy in treating diseases, and determining the optimal mechanical strength of the hydrogel in vivo is important for therapeutic success. We evaluated the proliferation of human neural stem cells incorporated within in situ-forming hydrogels and compared the effect of hydrogels with different elastic properties in cell/hydrogel-xenografted mice.

Methods: The gelatin-polyethylene glycol-tyramine (GPT) hydrogel was fabricated through enzyme-mediated cross-linking reaction using horseradish peroxidase (HRP) and hydrogen peroxide (H2O2).

Results: The F3-effluc encapsulated within a soft 1,800 pascal (Pa) hydrogel and stiff 5,800 Pa hydrogel proliferated vigorously in a 24-well plate until day 8. In vitro and in vivo kinetics of luciferase activity showed a slow time-to-peak after d-luciferin administration in the stiff hydrogel. When in vivo proliferation of F3-effluc was observed up to day 21 in both the hydrogel group and cell-only group, F3-effluc within the soft hydrogel proliferated more vigorously, compared to the cells within the stiff hydrogel. Ki-67-specific immunostaining revealed highly proliferative F3-effluc with compactly distributed cell population inside the 1,800 Pa or 5,800 Pa hydrogel.

Conclusions: We examined the in vivo effectiveness of different elastic types of hydrogels encapsulating viable neural stem cells by successfully monitoring the proliferation of implanted stem cells incorporated within a 3D hydrogel scaffold.

Original languageEnglish
Article number61
Pages (from-to)1-11
Number of pages11
JournalEJNMMI Research
Issue number1
StatePublished - 1 Jan 2014


  • Gelatin-based hydrogel
  • Human neural stem cell
  • In vivo bioluminescence imaging
  • Matrix elasticity
  • Optical kinetics

Cite this

Hwang, D. W., Park, K. M., Shim, H. K., Jin, Y., Oh, H. J., Oh, S. W., Lee, S., Youn, H., Joung, Y. K., Lee, H. J., Kim, S. U., Park, K. D., & Lee, D. S. (2014). In vivo bioluminescence imaging for viable human neural stem cells incorporated within in situ gelatin hydrogels. EJNMMI Research, 4(1), 1-11. [61].