title: Primary Cell Culture of Live Neurosurgically Resected Aged Adult Human Brain Cells and Single Cell Transcriptomics.
abstract: Investigation of human CNS disease and drug effects has been hampered by the lack of a system that enables single-cell analysis of live adult patient brain cells. We developed a culturing system, based on a papain-aided procedure, for resected adult human brain tissue removed during neurosurgery. We performed single-cell transcriptomics on over 300 cells, permitting identification of oligodendrocytes, microglia, neurons, endothelial cells, and astrocytes after 3 weeks in culture. Using deep sequencing, we detected over 12,000 expressed genes, including hundreds of cell-type-enriched mRNAs, lncRNAs and pri-miRNAs. We describe cell-type- and patient-specific transcriptional hierarchies. Single-cell transcriptomics on cultured live adult patient derived cells is a prime example of the promise of personalized precision medicine. Because these cells derive from subjects ranging in age into their sixties, this system permits human aging studies previously possible only in rodent systems.
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Spaethling
authors.affiliation: Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Young-Ji
authors.initials: YJ
authors.lastname: Na
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Jaehee
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Alexandra V
authors.initials: AV
authors.lastname: Ulyanova
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Gordon H
authors.initials: GH
authors.lastname: Baltuch
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Bell
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Brem
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: H Isaac
authors.initials: HI
authors.lastname: Chen
authors.affiliation: Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Hannah
authors.initials: H
authors.lastname: Dueck
authors.affiliation: Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Fisher
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Marcela P
authors.initials: MP
authors.lastname: Garcia
authors.affiliation: Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Mugdha
authors.initials: M
authors.lastname: Khaladkar
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: David K
authors.initials: DK
authors.lastname: Kung
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Timothy H
authors.initials: TH
authors.lastname: Lucas
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Donald M
authors.initials: DM
authors.lastname: O'Rourke
authors.affiliation: Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Derek
authors.initials: D
authors.lastname: Stefanik
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Jinhui
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Wolf
authors.affiliation: Department of Neurochemistry, Stockholm University, Stockholm 106 91, Sweden.
authors.forename: Tamas
authors.initials: T
authors.lastname: Bartfai
authors.affiliation: Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: M Sean
authors.initials: MS
authors.lastname: Grady
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Jai-Yoon
authors.initials: JY
authors.lastname: Sul
authors.affiliation: Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Junhyong
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: eberwine@upenn.edu.
authors.forename: James H
authors.initials: JH
authors.lastname: Eberwine
mesh_terms: D000328:Adult; D000368:Aged; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000378:metabolism; D017628:Microglia / Q000166:cytology / Q000378:metabolism; D008875:Middle Aged; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism; D025341:Principal Component Analysis; D062085:RNA, Long Noncoding / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D059010:Single-Cell Analysis; D059467:Transcriptome; D055815:Young Adult
chemical_list: D035683:MicroRNAs; D062085:RNA, Long Noncoding; D012333:RNA, Messenger
keywords: adult; aged; astrocyte; endothelial cell; human; microglia; neuron; primary cell; single cell; transcriptome