Spinal Cord Injection of hNSC or mT-mNSC

Four WPI, a second surgery was performed to transplant hNSC in Rag1 mice or mT-mNSC in CRYM-ZsGreen1 transgenic mice as previously described^{29 (link),31 (link),44 (link)}. After re-exposure of the spinal cord at C5, cells were injected using a NanoInjector with a micromanipulator (World Precision Instruments, Sarasota, FL) and siliconized beveled glass micropipettes (outer diameter = 100–110 mm; inner diameter = 70 mm; Sutter Instruments, Novato, CA). A total of 75,000 cells or vehicle (X-VIVO media) in 1 uL total volume was delivered to the spared spinal parenchyma at four sites, two rostral and two caudal to the lesion, 250 nL per site (schematic in Fig. 1c). All mice received postoperative care as described above.

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Nekanti U., Sakthivel P.S., Zahedi A., Creasman D.A., Nishi R.A., Dumont C.M., Piltti K.M., Guardamondo G.L., Hernandez N., Chen X., Song H., Lin X., Martinez J., On L., Lakatos A., Pawar K., David B.T., Guo Z., Seidlits S.K., Xu X., Shea L.D., Cummings B.J, & Anderson A.J. (2024). Multichannel bridges and NSC synergize to enhance axon regeneration, myelination, synaptic reconnection, and recovery after SCI. NPJ Regenerative Medicine, 9, 12.

Publication 2024

NanoInjector siliconized beveled glass micropipettes

Corresponding Organization : University of California, Irvine

Other organizations : University of Michigan–Ann Arbor, University of British Columbia, Rush University Medical Center, The University of Texas at Austin

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Variable analysis

independent variables

Type of cell transplanted (hNSC or mT-mNSC)

dependent variables

Survival and engraftment of transplanted cells in the spinal cord

control variables

Location of cell injection (four sites, two rostral and two caudal to the lesion)
Volume of cell/vehicle injection (1 uL total, 250 nL per site)
Injection method (NanoInjector with micromanipulator and siliconized beveled glass micropipettes)
Postoperative care

controls

Positive control: Transplantation of hNSC or mT-mNSC in Rag1 mice or CRYM-ZsGreen1 transgenic mice, respectively
Negative control: Injection of vehicle (X-VIVO media) instead of cells

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