BMPRⅡ^(+)neural precursor cells isolated and characterized from organotypic neurospheres:an in vitro model of human fetal spinal cord development  

作　　者：Michael W.Weible II Michael D.Lovelace Hamish D.Mundell Tsz Wai Rosita Pang Tailoi Chan-Ling 

机构地区：[1]Bosch Institute,Discipline of Anatomy and Histology(F13),University of Sydney,Sydney,NSW,Australia [2]School of Environment and Science,Griffith University,Nathan,QLD,Australia [3]Discipline of Medicine,Nepean Clinical School,Faculty of Medicine and Health,University of Sydney,Kingswood,NSW,Australia [4]New South Wales Brain Tissue Resource Centre,School of Medical Sciences,Faculty of Medicine and Health,University of Sydney,Charles Perkins Centre(D17),Sydney,NSW,Australia

出　　处：《Neural Regeneration Research》2024年第2期447-457,共11页中国神经再生研究（英文版）

基　　金：supported by grants from the National Health and Medical Research Council(NHMRC)of Australia(Nos.571100 and 1048082);the Baxter Charitable Foundation(to TCL);Medical Research grants from the Rebecca L.Cooper Medical Research Foundation(to MWW,TCL,and MDL);supported by a Charles D.Kelman,M.D.Postdoctoral Award(2010)from the International Retinal Research Foundation(USA)。

摘　　要：Roof plate secretion of bone morphogenetic proteins(BMPs)directs the cellular fate of sensory neurons during spinal cord development,including the formation of the ascending sensory columns,though their biology is not well understood.Type-ⅡBMP receptor(BMPRⅡ),the cognate receptor,is expressed by neural precursor cells during embryogenesis;however,an in vitro method of enriching BMPRⅡ^(+)human neural precursor cells(hNPCs)from the fetal spinal cord is absent.Immunofluorescence was undertaken on intact second-trimester human fetal spinal cord using antibodies to BMPRⅡand leukemia inhibitory factor(LIF).Regions of highest BMPRⅡ^(+)immunofluorescence localized to sensory columns.Parenchymal and meningeal-associated BMPRⅡ^(+)vascular cells were identified in both intact fetal spinal cord and cortex by co-positivity with vascular lineage markers,CD34/CD39.LIF immunostaining identified a population of somas concentrated in dorsal and ventral horn interneurons,mirroring the expression of LIF receptor/CD118.A combination of LIF supplementation and high-density culture maintained culture growth beyond 10 passages,while synergistically increasing the proportion of neurospheres with a stratified,cytoarchitecture.These neurospheres were characterized by BMPRⅡ^(+)/MAP2ab^(+/–)/βⅢ-tubulin^(+)/nestin^(–)/vimentin^(–)/GFAP^(–)/NeuN^(–)surface hNPCs surrounding a heterogeneous core ofβⅢ-tubulin^(+)/nestin^(+)/vimentin^(+)/GFAP^(+)/MAP2ab^(–)/NeuN^(–)multipotent precursors.Dissociated cultures from tripotential neurospheres contained neuronal(βⅢ-tubulin^(+)),astrocytic(GFAP+),and oligodendrocytic(O4+)lineage cells.Fluorescence-activated cell sorting-sorted BMPRⅡ^(+)hNPCs were MAP2ab^(+/–)/βⅢ-tubulin^(+)/GFAP^(–)/O4^(–)in culture.This is the first isolation of BMPRⅡ^(+)hNPCs identified and characterized in human fetal spinal cords.Our data show that LIF combines synergistically with high-density reaggregate cultures to support the organotypic reorganization of neurospheres,characteri

关 键 词：BMPRⅡ bone morphogenetic protein histotypic human spinal cord development leukemia inhibitory factor NEUROSPHERE ORGANOTYPIC reaggregate sensory columns 

分 类 号：R714.5[医药卫生—妇产科学]