Reclosure of surgically induced spinal open neural tube defects by the intraamniotic injection of human embryonic stem cells in chick embryos 24 hours after lesion induction

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Object

The authors previously reported that human embryonic stem cells (hESCs) injected into the amniotic cavity of chick embryos immediately after neural tube incision in a surgically induced spinal open neural tube defect (ONTD) model promote the reclosure capacity of neural tubes. To simulate more closely the clinical situation of human ONTDs, in which a ssubstantial time period elapses before the prenatal diagnosis of spinal ONTDs, the authors investigated whether this reclosure capacity remains enhanced by the intraamniotic injection of hESCs at 24 hours after ONTD induction.

Methods

One hundred twenty-two chick embryos with ONTDs were randomly assigned to two groups: the control group (59 embryos) and the hESC-injection group (hESC group, 63 embryos). After the neural tubes of both groups had been opened, the hESC group underwent direct intraamniotic injection with hESCs marked with an enhanced green fluorescent protein at 24 hours after ONTD induction. The lengths of the remnant ONTDs were measured and the presence of hESCs was determined at 4, 6, and 8 days after ONTD induction. No difference in survival rates was observed between the two groups. The mean length of the ONTDs, adjusted for body length at the time of death and initial lesion length, was significantly shorter in the hESC group than in the control group (p < 0.001). No hESCs were found within reclosed neural tubes; rather, they covered the defect area during the reclosure process.

Conclusions

The authors demonstrate that hESCs injected into the amniotic cavity at 24 hours after ONTD induction enhance reclosure ability in chick embryos.

Abbreviations used in this paper:CNS = central nervous system; eGFP = enhanced green fluorescent protein; GFAP = glial fibrillary acidic protein; hESC = human embryonic stem cell; NeuN = neuronal nuclei; ONTD = open neural tube defect; PBS = phosphate-buffered saline; POD = postoperative day.
Article Information

Contributor Notes

Address reprint requests to: Kyu-Chang Wang, M.D., Ph.D., Department of Neurosurgery, Seoul National University College of Medicine, 28 Yeongeon-dong, Jongno-gu, Seoul 110-744, Korea. email: kcwang@snu.ac.kr.Drs. D. H. Lee and E. Y. Kim contributed equally to this work.
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