Spinal Reflex Recovery after Dorsal Rhizotomy and Repair with Platelet-Rich Plasma (PRP) Gel Combined with Bioengineered Human Embryonic Stem Cells (hESCs)

Figure 1

(a–c) Cultivation of bioengineered human embryonic stem cell (hESC) clones over time. The formation of the cell monolayer (confluence) was observed on the fifth day (c). The cells were attached to a matrix (Matrigel) and expanded rapidly in their own medium, and they formed colonies and/or aggregates. Bright field. . (d) Clones were induced to overexpress FGF2 at 48 hours after doxycycline administration. Practically all cells were fluorescent green (GFP+), which indicates that hESCs were activated by DOX and then overexpressed FGF2. Phase contrast. . (e) A single bioengineered hESC was observed under electron microscopy, enabling visualization of the large nucleus, very evident nucleolus, and euchromatin, indicating high metabolic activity. . (f–h) Relative expression of basic fibroblast growth factor (FGF2), brain-derived neurotrophic factor (BDNF), and glial cell line-derived neurotrophic factor (GDNF) mRNA in vitro. (f) The transcript levels for FGF2 were over 35-fold higher when modified hESCs were induced by doxycycline. Modified hESCs not treated with doxycycline and wildtype cells treated or not treated with doxycycline presented nearly undetectable expression of FGF2. (g) Compared with other groups, the transcript levels for BDNF were significantly higher when modified hESCs were induced by doxycycline. (h) The transcript levels for GDNF were not significantly different between groups. . . hESCs: human embryonic stem cells; WT: wildtype; DOX: doxycycline.

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