Efficient differentiation of human embryonic stem cells into functional cerebellar-like cells.
The cerebellum has critical roles in motor and sensory learning and motor coordination. Many cerebellum-related disorders indicate cell therapy as a possible treatment of neural loss. Here we show that application of inductive signals involved in early patterning of the cerebellar region followed by application of different factors directs human embryonic stem cell differentiation into cerebellar-like cells such as granule neurons, Purkinje cells, interneuron, and glial cells. Neurons derived using our protocol showed a T-shaped polarity phenotype and express similar markers to the developed human cerebellum. Electrophysiological measurements confirmed functional electrical properties compatible with these cells. In vivo implantation of differentiated human embryonic stem cells transfected with MATH1-GFP construct into neonatal mice resulted in cell migration across the molecular and the Purkinje cell layers and settlement in the internal molecular layers. Our findings demonstrate that the universal mechanisms involved in the development of cerebellum can be efficiently recapitulated in vitro, which enables the design of new strategies for cell replacement therapy, to study early human development and pathogenesis of neurodegenerative diseases.
Erceg
Slaven
S
Cellular Reprogramming Laboratory, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain. serceg@cipf.es
Ronaghi
Mohammad
M
Zipancic
Ivan
I
Lainez
Sergio
S
Roselló
Mireia Gárcia
MG
Xiong
Chen
C
Moreno-Manzano
Victoria
V
Rodríguez-Jiménez
Fernando Javier
FJ
Planells
Rosa
R
Alvarez-Dolado
Manuel
M
Bhattacharya
Shom Shanker
SS
Stojkovic
Miodrag
M
D000200:Action Potentials; D000328:Adult; D000368:Aged; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002465:Cell Movement; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D005260:Female; D005333:Fetus / Q000033:anatomy & histology; D005865:Gestational Age; D006801:Humans; D008297:Male; D051379:Mice; D008875:Middle Aged; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011247:Pregnancy; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D033581:Stem Cell Transplantation; D055815:Young Adult
D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D011993:Recombinant Fusion Proteins; C500942:ATOH1 protein, human