Human pluripotent stem cells-derived inner ear organoids recapitulate otic development in vitro.

Our molecular understanding of the early stages of human inner ear development has been limited by the difficulty in accessing fetal samples at early gestational stages. As an alternative, previous studies have shown that inner ear morphogenesis can be partially recapitulated using induced pluripotent stem cells (iPSCs) directed to differentiate into Inner Ear Organoids (IEOs). Once validated and benchmarked, these systems could represent unique tools to complement and refine our understanding of human otic differentiation and model developmental defects. Here, we provide the first direct comparisons of the early human embryonic otocyst and human iPSC-derived IEOs. We use multiplexed immunostaining, and single-cell RNA sequencing to characterize IEOs at three key developmental steps, providing a new and unique signature of in vitro derived otic -placode, -epithelium, -neuroblasts, and -sensory epithelia. In parallel, we evaluate the expression and localization of critical markers at these equivalent stages in human embryos. We show that the placode derived in vitro (days 8-12) has similar marker expression to the developing otic placode of Carnegie Stage (CS) 11 embryos and subsequently (days 20-40) this gives rise to otic epithelia and neuroblasts comparable to the CS13 embryonic stage. Differentiation of sensory epithelia, including supporting cells and hair cells starts in vitro at days 50-60 of culture. The maturity of these cells is equivalent to vestibular sensory epithelia at week 10 or cochlear tissue at week 12 of development, before functional onset. Together, our data indicate that the current state-of-the-art protocol enables the specification of bona fide otic tissue, supporting the further application of IEOs to inform inner ear biology and disease.

Functional muscle strength recovery from nail gun injury to the primary motor cortex.

Aim: Functional recovery following injury to the primary motor cortex is an uncommon phenomenon, given the limited ability of neurons of the adult central nervous system to regenerate. Case description: We report on a patient with near complete functional muscle strength recovery from a marked monoparesis due to nail gun injury to the medial primary motor cortex. Besides surgical decision-making, we discuss possible related mechanisms and current challenges in the regenerative processes responsible for the functional recovery. Discussion: To achieve a favorable outcome, surgical decision-making to prevent secondary damage is of upmost importance. Lesion-induced inflammatory response may potentiate endogenous neurogenesis and neuronal plasticity and potentially contribute to the regenerative process involved.

Endothelial Progenitor Cell-Derived Factors Exert Neuroprotection in Cultured Cortical Neuronal Progenitor Cells.

There is substantial evidence that stem and progenitor cells secrete trophic factors that have potential for repairing injured tissues. We have previously reported that the conditioned medium (CM) obtained from endothelial progenitor cells (EPC) cultures protects striatal neurons against 3-nitropropionic acid-induced toxicity. In the present study we tested the hypothesis that EPC-CM may support cortical neuronal cell function and/or survival. EPC were isolated from the peripheral blood of healthy human donors and cultured in hypoxic conditions (1.5% O2) to stimulate the secretion of growth factors. The supernatant or conditioned medium (EPC-CM) was then collected and used for the various experiments. Primary cultures of cerebral cortex from fetal rat embryonic day 14 were treated with EPC-CM and challenged by glucose and serum deprivation. We observed that EPC-CM treatment significantly increased total cell number and cell viability in the cultures. Similarly, the number of lba1-expressing cells was significantly upregulated by EPC-CM, while western blot analyses for the astroglial marker glial fibrillary acidic protein did not show a marked difference. Importantly, the number of beta-lll-tubulin-positive neurons in the cultures was significantly augmented after EPC-CM treatment. Similarly, western blot analyses for beta-III-tubulin showed significant higher signal intensities. Furthermore, EPC-CM administration protected neurons against glucose- and serum deprivation-induced cell loss. In sum, our findings identified EPC-CM as a means to promote viability and/or differentiation of cortical neurons and suggest that EPC-CM might be useful for neurorestorative approaches.

Endothelial Progenitor Cells Conditioned Medium Supports Number of GABAergic Neurons and Exerts Neuroprotection in Cultured Striatal Neuronal Progenitor Cells.

There is growing evidence that stem and progenitor cells exert regenerative actions by means of paracrine factors. In line with these notions, we recently demonstrated that endothelial progenitor cell (EPC)-derived conditioned medium (EPC-CM) substantially increased viability of brain microvascular cells. In the present study, we aimed at investigating whether EPC-CM supports cell survival of cultured striatal progenitor cells. For that purpose, primary cultures from fetal rat embryonic (E14) ganglionic eminence were prepared and grown for 7 days in vitro (DIV). EPC-CM was administered from DIV5-7. Treatment of the striatal cultures with EPC-CM resulted in significantly increased densities of GABA-immunoreactive (-ir) neurons. Inhibition of mitogen-activated protein kinase and phosphatidylinositol-3-kinase, but not of the ROCK pathway, significantly attenuated the EPC-CM induced increase in GABA-ir cell densities. Similar results were observed when EPC-CM was subjected to proteolytic digestion and lipid extraction. Furthermore, inhibition of translation abolished the EPC-CM induced effects. Importantly, EPC-CM displayed neuroprotection against 3-nitropropionic acid induced toxicity. These findings demonstrate that EPC-derived paracrine factors substantially promote survival and/or differentiation of cultured striatal progenitor cells involving both proteinaceous factors and lipidic factors. In sum, EPC-CM constituents might lead to a novel cell-free therapeutic strategy to challenge neuronal degeneration.

Cell-based therapy facilitates venous thrombus resolution.

Endothelial progenitor cells (EPC) are involved in many healing processes in cardiovascular diseases and can be found in spontaneously resolving venous thrombi. The purpose of the present study was to investigate whether the therapeutic administration of EPC might enhance the resolution of venous thrombi. For this purpose, venous thrombosis was induced in the infrarenal inferior vena cava (IVC) in 28 athymic nude rats. Culture expanded EPC derived from human peripheral blood mononuclear cells were injected intravenously two and four days after thrombus induction. Recanalisation of the IVC and thrombus organisation were assessed by laser Doppler measurements of the blood flow and immunohistochemical detection of endothelialised luminal structures in the thrombus. EPC transplantation resulted in significantly enhanced thrombus neovascularisation (capillary density: 186.6 +/- 26.7/HPF vs. 78 +/- 12.3/HPF, p<0.01; area covered by capillaries: 8.9 +/- 1.7 microm(2) vs. 2.5 +/- 1.3 microm(2), p<0.01) and was accompanied by a substantial increase in intra-thrombus blood flow (perfusion ratio: 0.7 +/- 0.07 vs. 0.3 +/- 0.08, p<0.02). These results were paralleled by augmented macrophage recruitment into resolving thrombi in the animals treated with EPC (39.4 +/- 4.7/HPF vs. 11.6 +/- 1.9/HPF, p<0.01). Our data suggest that EPC transplantation might be of clinical value to facilitate venous thrombus resolution in cases where current therapeutic options have limited success.