Obtaining Human Microglia from Adult Human Brain Tissue

Author(s):  
Ishan Agrawal ◽  
Shivanjali Saxena ◽  
Preethika Nair ◽  
Deepak Jha ◽  
Sushmita Jha
2020 ◽  
Vol 123 (3) ◽  
pp. 945-965 ◽  
Author(s):  
Kevin Lee ◽  
Thomas I.-H. Park ◽  
Peter Heppner ◽  
Patrick Schweder ◽  
Edward W. Mee ◽  
...  

The human brain shows remarkable complexity in its cellular makeup and function, which are distinct from nonhuman species, signifying the need for human-based research platforms for the study of human cellular neurophysiology and neuropathology. However, the use of adult human brain tissue for research purposes is hampered by technical, methodological, and accessibility challenges. One of the major problems is the limited number of in vitro systems that, in contrast, are readily available from rodent brain tissue. With recent advances in the optimization of protocols for adult human brain preparations, there is a significant opportunity for neuroscientists to validate their findings in human-based systems. This review addresses the methodological aspects, advantages, and disadvantages of human neuron in vitro systems, focusing on the unique properties of human neurons and synapses in neocortical microcircuits. These in vitro models provide the incomparable advantage of being a direct representation of the neurons that have formed part of the human brain until the point of recording, which cannot be replicated by animal models nor human stem-cell systems. Important distinct cellular mechanisms are observed in human neurons that may underlie the higher order cognitive abilities of the human brain. The use of human brain tissue in neuroscience research also raises important ethical, diversity, and control tissue limitations that need to be considered. Undoubtedly however, these human neuron systems provide critical information to increase the potential of translation of treatments from the laboratory to the clinic in a way animal models are failing to provide.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sven Hildebrand ◽  
Anna Schueth ◽  
Klaus von Wangenheim ◽  
Christian Mattheyer ◽  
Francesco Pampaloni ◽  
...  

Biologia ◽  
2009 ◽  
Vol 64 (4) ◽  
Author(s):  
Ivana Macikova ◽  
Anna Perzelova ◽  
Peter Mraz ◽  
Ivan Bizik ◽  
Juraj Steno

AbstractTraditionally, astrocytes are divided into fibrous and protoplasmic types based on their morphologic appearance. Here the cultures were prepared separately from the adult human cortical gray and white matter of brain biopsies. Both cultures differed only in the number of glial fibrillary acidic protein (GFAP)-positive cells. In the gray matter these were absent or rare, whereas in confluent cultures from the white matter they reached 0.1% of all cells. Three main morphologic types of GFAP-positive cells were found in this study: stellate, bipolar and large flat cells. GFAP-positive cells with two or three long processes mimic a neuron-like morphology. We did not find process-bearing cells expressing neuronal markers (MAP-2, NF, and N-CAM). The conflicting reports concerning GFAP immunostaining and the study dealing with the presence of putative neurons in adult human brain cultures are discussed with respect to these findings. The latter classification of astrocytes into type 1 and type 2 is based on immunostaining to A2B5 antigen: type 1 (GFAP+/A2B5−) and type 2 (GFAP+/A2B5+) astrocytes are proposed to be analogous to protoplasmic and fibrous astrocytes, respectively. In adult human brain cultures we found only small amount of A2B5-positive cells. Double immunofluorescence revealed that astroglial cells of similar fibrous or bipolar shape grown on one coverslip were either GFAP+/A2B5+ or GFAP+/A2B5−. On the other hand, the A2B5+/GFAP− immunophenotype was not observed. These results indicate that in general the cell phenotype from adult human brain tissue is not well established when they are in culture.


2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Claudia P. Figueiredo ◽  
Fernanda G. Q. Barros-Aragão ◽  
Rômulo L. S. Neris ◽  
Paula S. Frost ◽  
Carolina Soares ◽  
...  

Cryobiology ◽  
2003 ◽  
Vol 47 (2) ◽  
pp. 179-183 ◽  
Author(s):  
Jean-François Brunet ◽  
Luc Pellerin ◽  
Pierre Magistretti ◽  
Jean-Guy Villemure

Biologia ◽  
2007 ◽  
Vol 62 (5) ◽  
Author(s):  
Anna Perzelova ◽  
Ivana Macikova ◽  
Marcienne Tardy ◽  
Peter Mraz ◽  
Ivan Bizik ◽  
...  

AbstractGlial fibrillary acidic protein (GFAP) is an intermediate filament protein considered to be the best astroglial marker. However, the predominant cell population in adult human brain tissue cultures does not express GFAP; these cells have been termed “glia-like” cells. The basic question about histological origin of adult human brain cultures remains unanswered. Some authors showed that “glia-like” cells in adult human brain cultures might be of non-glial origin. We examined primary explant tissue cultures derived from 70 adult human brain biopsies. Within first 5–10 days approximately 5–10% of the small explants became attached. Outgrowing cells were mostly flat cells. These cells formed confluent layer over 3–6 weeks in culture. At confluence the cultures contained 2–5% of microglial cells, 0.1% GFAP-positive astrocytes, less than 0.01% oligodendrocytes and 95–98% GFAP-negative “glia-like” cells. This population of flat “glia-like” cells was positively stained for vimentin, fibronectin, and 20–30% of these cells stained for nestin. Our findings revealed that 1 mM dibutyryl-cAMP addition, in serum free conditions, induced a reversible stellation in 5-10% of the flat “glia-like” cells but did not induce the expression of GFAP or nestin in morphologically changed stellate cells. These results demonstrate that “glia-like” cells in primary adult human brain cultures constitute heterogeneous cell populations albeit with similar morphological features. Two distinct subpopulations have been shown: (i) the one immunostained for nestin; and (ii) the other reactive for dibutyryl-cAMP treatment.


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