Arctic ground squirrel neuronal progenitor cells resist oxygen and glucose deprivation-induced death

• 作者: Austin P Ross Judith Kelleher-Andersson Kelly L Drew Matthew Wells Rebecca McGee
• 来源期刊: 世界生物化学杂志：英文版（电子版）
• 学科: 医学
• 关键词: NEUROGENESIS NEURONAL PROGENITOR Hypoxia tolerance HIBERNATION
• 年，卷（期）: 2016,（1）
• 页码范围: 168-177
• 页数: 10页
• 分类号: R741

摘要

AIM: To investigate the influence of ischemia/reperfusion on arctic ground squirrel（AGS） neuronal progenitor cells（NPCs）, we subjected these cultured cells to oxygen and glucose deprivation.METHODS: AGS NPCs were expanded and differentiated into NPCs and as an ischemia vulnerable control, commercially available human NPCs（hNPCs） were seeded from thawed NPCs. NPCs, identified by expression of TUJ1 were seen at 14-21 d in vitro（DIV）. Cultures were exposed to control conditions, hypoxia, oxygen and glucose deprivation or glucose deprivation alone or following return to normal conditions to model reperfusion. Cell viability and death were assessed from loss of ATP as well as from measures of alamarB lue and lactate dehydrogenase in the media and from counts of TUJ1 positive cells using immunocytochemistry. Dividing cells were identified by expression of Ki67 and phenotyped by double labeling with GFAP, MAP2 ab or TUJ1. RESULTS: We report that when cultured in NeuraLifeTM, AGS cells remain viable out to 21 DIV, continue to express TUJ1 and begin to express MAP2 ab. Viability of hN PCs assessed by fluorescence alamarB lue（arbitrary units） depends on both glucose and oxygen availability [viability of hNPCs after 24 h oxygen glucose deprivation（OGD） with return of oxygen and glucose decreased from 48151 ± 4551 in control cultures to 43481 ± 2413 after OGD, P < 0.05]. By contrast, when AGS NPCs are exposed to the same OGD with reperfusion at 14 DIV, cell viability assessed by alamar Blue increased from 165305 ± 11719 in control cultures to 196054 ± 13977 after OGD. Likewise AGS NPCs recovered ATP（92766 ± 6089 in control and 92907 ± 4290 after modeled reperfusion; arbitrary luminescence units）, and doubled in the ratio of TUJ1 expressing neurons to total dividing cells（0.11 ± 0.04 in control cultures vs 0.22 ± 0.2 after modeled reperfusion, P < 0.05）. Maintaining AGS NPCs for a longer time in culture lowered resistance to injury, however, did not impair proliferation of NPCs relat