In Vivo Bioluminescence Imaging for Prolonged Survival of Transplanted Human Neural Stem Cells Using 3D Biocompatible Scaffold in Corticectomized Rat Model

Abstract

Stem cell-based treatment of traumatic brain injury has been limited in its capacity to bring about complete functional recovery, because of the poor survival rate of the implanted stem cells. It is known that biocompatible biomaterials play a critical role in enhancing survival and proliferation of transplanted stem cells via provision of mechanical support. In this study, we noninvasively monitored in vivo behavior of implanted neural stem cells embedded within poly-l-lactic acid (PLLA) scaffold, and showed that they survived over prolonged periods in corticectomized rat model. Corticectomized rat models were established by motor-cortex ablation of the rat. F3 cells expressing enhanced firefly luciferase (F3-effLuc) were established through retroviral infection. The F3-effLuc within PLLA was monitored using IVIS-100 imaging system 7 days after corticectomized surgery. F3-effLuc within PLLA robustly adhered, and gradually increased luciferase signals of F3-effLuc within PLLA were detected in a day dependent manner. The implantation of F3-effLuc cells/PLLA complex into corticectomized rats showed longer-lasting luciferase activity than F3-effLuc cells alone. The bioluminescence signals from the PLLA-encapsulated cells were maintained for 14 days, compared with 8 days for the non-encapsulated cells. Immunostaining results revealed expression of the early neuronal marker, Tuj-1, in PLLA-F3-effLuc cells in the motor-cortex-ablated area. We observed noninvasively that the mechanical support by PLLA scaffold increased the survival of implanted neural stem cells in the corticectomized rat. The image-guided approach easily proved that scaffolds could provide supportive effect to implanted cells, increasing their viability in terms of enhancing therapeutic efficacy of stem-cell therapy.

Human neural stem cell intracerebral grafts show spontaneous early neuronal differentiation after several weeks

“…First studies showed a positive outcome using bioluminescence reporters for neuroimaging [34,35]. In the present study optical cell tracking with Luc2 was highly efficient.…”

miR-192 suppresses leptomeningeal dissemination of medulloblastoma by modulating cell proliferation and anchoring through the regulation ofDHFR, integrins, andCD47

“…To visualize the transplanted D283 cells, D283 cells were transfected with an effLuc viral vector and a DNA vector carrying major structural proteins (GAG, Pol, and Env) using lipofectamine 2000 (Invitrogen) as described previously [ 43 ]. The infected cells were sorted by magnetic-activated cell sorting (Miltenyi Biotech Ltd.) through monoclonal anti-CD90.1 conjugated to magnetic microbeads.…”

Stem Cell Imaging: Tools to Improve Cell Delivery and Viability

“…For example, the decellularized porcine ventricular ECM can effectively support cardiac progenitor cells by showing a strong serum-induced proliferation and a resistance to apoptosis in serum starvation [ 134 ]; the poly(ethylene glycol) diacrylate (PEGDA) hydrogel stimulates the implanted MSCs in cartilage repair and causes less pain to the patients [ 135 ]. Poly-L-lactic acid (PLLA) scaffolds can prolong the survival and increase the proliferation rate of F3 cells implanted to ablated motor cortex of the corticectomized rat [ 136 ]. Taking advantages of these extra support structures can improve SCT and increase cell viability.…”