Healthy human CSF promotes glial differentiation of hESC-derived neural cells while retaining spontaneous activity in existing neuronal networks

Abstract

SummaryThe possibilities of human pluripotent stem cell-derived neural cells from the basic research tool to a treatment option in regenerative medicine have been well recognized. These cells also offer an interesting tool for in vitro models of neuronal networks to be used for drug screening and neurotoxicological studies and for patient/disease specific in vitro models. Here, as aiming to develop a reductionistic in vitro human neuronal network model, we tested whether human embryonic stem cell (hESC)-derived neural cells could be cultured in human cerebrospinal fluid (CSF) in order to better mimic the in vivo conditions. Our results showed that CSF altered the differentiation of hESC-derived neural cells towards glial cells at the expense of neuronal differentiation. The proliferation rate was reduced in CSF cultures. However, even though the use of CSF as the culture medium altered the glial vs. neuronal differentiation rate, the pre-existing spontaneous activity of the neuronal networks persisted throughout the study. These results suggest that it is possible to develop fully human cell and culture-based environments that can further be modified for various in vitro modeling purposes.

Human Cerebrospinal Fluid Regulates Proliferation and Migration of Stem Cells Through Insulin-Like Growth Factor-1

“…The artificial CSF was prepared as follows: H 2 O, NaCl (124 mM), KCl (2.5 mM), NaH 2 PO 4 (1.25 mM), CaCl 2 $2H 2 O (2.5 mM), MgSO 4 $7H 2 O (1.5 mM), and glucose (10 mM). Then, NaHCO 3 was used to adjust the pH during 7.35-7.45 [25,26].…”

Recommendation on test readiness criteria for new approach methods in toxicology: Exemplified for developmental neurotoxicity

“…Recently, synapsin as a pre-synaptic vesicle protein was detected by staining in hiPSC-derived mixed cultures that contain GABAergic, glutamatergic and dopaminergic neurons (Zagoura et al, 2017), however, neither synapse number nor protein expression were quantified. Functionality of synapses in these cultures was displayed by electrical activity on microelectrode arrays (MEA), i.e., spikes and bursts, but do not seem to present synchronized bursting as seen for rat primary cortical culture-derived networks (Brown et al, 2016(Brown et al, , 2017 or hESC-derived cultures on MEA chips (Kapucu et al, 2012;Kiiski et al, 2013). Nonetheless, MEA measurements were already successfully applied for in vitro DNT testing during chronic exposure to domoic acid , including evaluation of different receptor subtype involvement (Hogberg and Bal-Price, 2011), MeHg ical features of rosette formation or changes in gene expression levels below the cytotoxic threshold.…”

Effect of Multiple Sclerosis Cerebrospinal Fluid and Oligodendroglia Cell Line Environment on Human Wharton’s Jelly Mesenchymal Stem Cells Secretome

“…This study was designed to assess the immunomodulatory potential of WJ-MSCs, alone and in contact with adult human CSF derived from MS patients and healthy controls. So far, the observations of the interaction between CSF and neural stem cells have been successfully conducted with cells of embryonic [15,26,27] or of adult origin (BM-MSCs) [28,29]. To our knowledge, this is the first study with the use of human WJ-MSCs and adult human CSFs collected from MS patients as well as healthy controls, assessing the humoral response in such a wide spectrum.…”