Improved methods for the differentiation of hypothalamic vasopressin neurons using mouse induced pluripotent stem cells

Rostral Hypothalamic Differentiation With Minimal Exogenous Signals in Human Naive Induced Pluripotent Stem Cells

“…Our previous study using miPSCs showed that stopping miPSC colonies to clumps of approximately 10-20 cells, rather than isolating them to a single cell for SFEBq, can facilitate miPSC aggregation 29 . Accordingly, SFEBq using clumps was performed to prevent collapse and cyst formation.…”

“…Using a ROCK inhibitor, primed hESCs/hiPSCs can be differentiated into non-rostral nervous systems such as the telencephalon 17,18 , cerebellum 22 , and spinal cord 24 . However, for inducing the rostral part of the central nervous system, such as AVP neurons, additional conditions are required to induce the rostral edge of the neural plate by using minimal exogenous signals as the positional information during differentiation 28,29 . In the primed hESC/hiPSC cultures, use of the ROCK inhibitor alone cannot maintain their aggregation in the state of minimal exogenous signals.…”

“…Pluripotent stem cells can differentiate into various cells or tissues in three-dimensional cultures termed "serum-free culture of embryoid body-like aggregates with quick re-aggregation" (SFEBq) [17][18][19][20][21][22][23][24][25][26][27] . Based on the differentiation of hypothalamic neurons from mouse embryonic stem cells (mESCs) by SFEBq 28 , we improved the differentiation e ciency of mouse iPSCs 29 . In these methods, the differentiation of pluripotent stem cells into neural tissue is achieved by regulating the positional information added to the medium.…”

“…S1A). To differentiate the hypothalamus from mouse ES/iPSCs, it is important to remove exogenous signals strictly in the early stages of differentiation, and to control the positional information so that it represents the most rostral side 28,29 . The medium with minimized exogenous signals is called a growth factor-free chemically de ned medium (gfCDM) 28 .…”

Hypothalamic–pituitary organoid generation through the recapitulation of organogenesis

“…We improved the differentiating methods of hypothalamic AVP neurons with mouse iPS cells (Mitsumoto et al., 2019). Three‐dimensional floating culture was also effective for hypothalamic differentiation from mouse iPS cells.…”