Direct isolation and RNA-seq reveal environment-dependent properties of engrafted neural stem/progenitor cells.
Neural stem/progenitor cell (NSPC) transplantation is a promising treatment for various neurodegenerative disorders including spinal cord injury, however, no direct analysis has ever been performed on their in vivo profile after transplantation. Here we combined bioimaging, flow-cytometric isolation and ultra-high-throughput RNA sequencing to evaluate the cellular properties of engrafted NSPCs. The acutely transplanted NSPCs had beneficial effects on spinal cord injury, particularly neuroprotection and neurohumoral secretion, whereas their in situ secretory activity differed significantly from that predicted in vitro. The RNA-sequencing of engrafted NSPCs revealed dynamic expression/splicing changes in various genes involved in cellular functions and tumour development depending on graft environments. Notably, in the pathological environment, overall transcriptional activity, external signal transduction and neural differentiation of engrafted NSPCs were significantly suppressed. These results highlight the vulnerability of engrafted NSPCs to environmental force, while emphasizing the importance of in situ analysis in advancing the efficacy and safety of stem cell-based therapies.
Department of Orthopedic Surgery, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Hiromi
H
Kumamaru
Yasuyuki
Y
Ohkawa
Hirokazu
H
Saiwai
Hisakata
H
Yamada
Kensuke
K
Kubota
Kazu
K
Kobayakawa
Koichi
K
Akashi
Hideyuki
H
Okano
Yukihide
Y
Iwamoto
Seiji
S
Okada
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000235:genetics; D060833:Cellular Microenvironment / Q000235:genetics; D016543:Central Nervous System Neoplasms / Q000235:genetics / Q000473:pathology; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D007249:Inflammation / Q000473:pathology; D008163:Luminescent Measurements; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000378:metabolism; D020127:Recovery of Function; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000235:genetics; D013116:Spinal Cord / Q000473:pathology / Q000503:physiopathology; D013119:Spinal Cord Injuries / Q000473:pathology / Q000503:physiopathology / Q000628:therapy; D033581:Stem Cell Transplantation; D059467:Transcriptome / Q000235:genetics
D018696:Neuroprotective Agents