Production of a Bioink Containing Decellularized Spinal Cord Tissue for 3D Bioprinting.

For the past few years, three-dimensional (3D) bioprinting has emerged as a promising approach in the field of regenerative medicine. This technique allows for the production of 3D scaffolds to support cell transplantation due to its ability to mimic the extracellular environment. One alternative to enhancing cell adhesion, survival, and proliferation is the use of decellularized extracellular matrix as a bioink component. The aim of this study was to produce a bioink using lyophilized rat decellularized spinal cord tissue (DSCT) for 3D bioprinting of nervous tissue. DNA quantification, hematoxylin and eosin and DAPI staining indicated that 1% sodium dodecyl sulfate and 9 h processing were effective in removing the cells from the spinal cord samples. The cell viability assay showed that the decellularized matrix is not cytotoxic for PC12 cells. The hydrogel containing DSCT, alginate, and gelatine used as the base for the bioink has a shear thinning behavior and low G″/G' ratio, allowing for good printability without compromising cell viability after 3D bioprinting. The bioink supported long-term PC12 cell survival, with 93% of live cells 4 weeks after printing, and stimulated the production of laminin-1 and neurofilament-M. This bioink, therefore, represents an easily available biomaterial for central nervous system tissue engineering.

Secretome of stem cells from human exfoliated deciduous teeth (SHED) and its extracellular vesicles improves keratinocytes migration, viability, and attenuation of H2 O2 -induced cytotoxicity.

Therapies for wound healing using the secretome and extracellular vesicles (EVs) of mesenchymal stem/stromal cells have been shown to be successful in preclinical studies. This study aimed to characterise the protein content of the secretome from stem cells from human exfoliated deciduous teeth (SHED) and analyse the in vitro effects of SHED-conditioned medium (SHED-CM) and SHED extracellular vesicles (SHED-EVs) on keratinocytes. EVs were isolated and characterised. The keratinocyte viability and migration of cells treated with SHED-EVs and conditioned medium (CM) were evaluated. An HaCaT apoptosis model induced by H2 O2 in vitro was performed with H2 O2 followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and live/dead assays. Finally, the expression of vascular endothelial growth factor (VEGF) in keratinocytes treated with secretome and EVs was evaluated by immunofluorescence staining and confirmed with RT-qPCR. SHED-EVs revealed a cup-shaped morphology with expression of the classical markers for exosomes CD9 and CD63, and a diameter of 181 ± 87 nm. The internalisation of EVs by HaCaT cells was confirmed by fluorescence microscopy. Proteomic analysis identified that SHED-CM is enriched with proteins related to stress response and development, including cytokines (CXCL8, IL-6, CSF1, CCL2) and growth factors (IGF2, MYDGF, PDGF). The results also indicated that 50% CM and 0.4-0.6 µg/mL EVs were similarly efficient for improving keratinocyte viability, migration, and attenuation of H2 O2 -induced cytotoxicity. Additionally, expression of VEGF on keratinocytes increased when treated with SHED secretome and EVs. Furthermore, VEGF gene expression in keratinocytes increased significantly when treated with SHED secretome and EVs. Both SHED-CM and SHED-EVs may therefore be promising therapeutic tools for accelerating re-epithelialization in wound healing.

Cell Electrospinning: a Review of Materials and Methodologies for Biofabrication.

The process of electrohydrodynamic living cell microencapsulation inside a scaffold during the electrospinning (ES) process is called cell electrospinning (CE). Several studies demonstrate the feasibility of using cell electrospinning for biomedical applications, allowing for the direct biofabrication of living cells to be encapsulated in fibers for the formation of active biological scaffolds. In this review, a comprehensive overview of the materials and methodologies used in cell electrospinning, as well as their biomedical application in tissue engineering, is provided. Cell ES represents an innovative technique for automated application in regenerative medicine.

Easy-to-Assembly System for Decellularization and Recellularization of Liver Grafts in a Bioreactor.

Decellularization of organs creates an acellular scaffold, ideal for being repopulated by cells. In this work, a low-cost perfusion system was created to be used in the process of liver decellularization and as a bioreactor after recellularization. It consists of a glass chamber to house the organ coupled to a peristaltic pump to promote liquid flow through the organ vascular tree. The rats' liver decellularization was made with a solution of sodium dodecyl sulfate. The recellularization was made with 108 mesenchymal stromal/stem cells and cultivated for seven days. The decellularized matrices showed an absence of DNA while preserving the collagen and glycosaminoglycans quantities, confirming the efficiency of the process. The functional analyses showed a rise in lactate dehydrogenase levels occurring in the first days of the cultivation, suggesting that there is cell death in this period, which stabilized on the seventh day. Histological analysis showed conservation of the collagen web and some groups of cells next to the vessels. It was possible to establish a system for decellularization and a bioreactor to use for the recellularization method. It is easy to assemble, can be ready to use in little time and be easily sterilized.

Bilayer scaffold from PLGA/fibrin electrospun membrane and fibrin hydrogel layer supports wound healingin vivo.

Hybrid scaffolds from natural and synthetic polymers have been widely used due to the complementary nature of their physical and biological properties. The aim of the present study, therefore, has been to analyzein vivoa bilayer scaffold of poly(lactide-co-glycolide)/fibrin electrospun membrane and fibrin hydrogel layer on a rat skin model. Fibroblasts were cultivated in the fibrin hydrogel layer and keratinocytes on the electrospun membrane to generate a skin substitute. The scaffolds without and with cells were tested in a full-thickness wound model in Wistar Kyoto rats. The histological results demonstrated that the scaffolds induced granulation tissue growth, collagen deposition and epithelial tissue remodeling. The wound-healing markers showed no difference in scaffolds when compared with the positive control. Activities of antioxidant enzymes were decreased concerning the positive and negative control. The findings suggest that the scaffolds contributed to the granulation tissue formation and the early collagen deposition, maintaining an anti-inflammatory microenvironment.

Production of nanostructured systems: Main and innovative techniques.

In the constant search for the development of more-specific and more-selective drugs, especially with regard to the challenge of encapsulating hydrophilic molecules, polymer nanotechnologies are remarkable for their biocompatible and biodegradable properties. The most-used nanoencapsulation methods consist of emulsification procedures, where emulsified droplets of a given polymer and drug solidify into nanoparticles after solvent extraction from the polymeric phase. This review introduces conventional emulsification methods but also highlights new emulsification technologies such as microfluidics, membrane emulsification and other techniques, including spray drying, inkjet printing and electrospraying.

Avaliação da composição química de um óleo de copaíba (Copaifera spp) e seu efeito em células-tronco mesenquimais / Evaluation of the chemical composition of a copaiba oil (Copaifera spp) and its effect on mesenchymal stem cells

A oleorresina obtida de copaíferas é amplamente utilizada na medicina tradicional brasileira. Este estudo avaliou a composição química por Cromatografia Gasosa (CG) e o efeito da oleorresina de Copaifera officinalis em células-tronco. Para isso as células foram tratadas com a oleorresina nas concentrações de 0,5, 20, 110, 140, 170 ou 200 µg/ml por 24h. A avaliação por CG identificou os sesquiterpenos ß-cariofileno, trans-α-bergamoteno e óxido de cariofileno II como os compostos majoritários da oleorresina. As menores concentrações de oleorresina utilizadas apresentaram resultados semelhantes ao grupo controle e as maiores concentrações diminuíram significativamente a viabilidade celular e apresentaram maior citotoxicidade. Como conclusão, os principais componentes encontrados na oleorresina de copaíba foram os sesquiterpenos e as baixas concentrações testadas não foram citotóxicas. O aumento das concentrações de oleorresina de copaíba promoveu diminuição da viabilidade celular e aumento dos efeitos citotóxicos nas células-tronco. Embora a oleorresina de copaíba tenha uso etnofarmacológico na cicatrização, este estudo demonstrou efeito citotóxico em células-tronco, as quais estão relacionadas ao processo de regeneração corpóreo. Portanto, deve-se ter cuidado com a dosagem de oleorresina a ser utilizada, uma vez que este estudo in vitro mostrou citotoxicidade e um impacto negativo na viabilidade das células-tronco nas mais altas concentrações testadas. [au]

Oleoresinobtained from Copaifera trees is extensively used in Brazilian traditional medicine. This study hasevaluated the chemical composition and effect of Copaifera officinalisoleoresin on stem cells. The oleoresin was analyzed by Gas Chromatography (GC) and the cells were treated with the oleoresin at concentrations of 0.5, 20, 110, 140, 170 or 200 µg/ml for 24h for cellular tests. GC identified the sesquiterpenes beta-caryophyllene, trans-alpha-bergamotene, and caryophyllene oxide II as the main compounds in oleoresin. The cell viability and cytotoxicity assays showed the lowest concentrations of oleoresin used presented similar results to the control group and the higher concentrations tested significantly decreased cell viability and increased cytotoxicity. In a conclusion, the main components found in copaiba oleoresin were sesquiterpenes and the low tested concentrations were not cytotoxic. The increased concentrations of copaiba oleoresin promoted a decrease in cell viability and an increase of cytotoxicity in the stem cells. Although copaiba oleoresin has ethnopharmacology use in healing, this study showed toxicity in stem cells, which are related to the corporeal regeneration process. Therefore, caution must be taken with the dosage of the oleoresin to be used since this in vitro study showed cytotoxicity and a negative impact on stem cell viability at the higher tested concentrations. [au]

Toxicity of oleate-based amino protic ionic liquids towards Escherichia coli, Danio rerio embryos and human skin cells.

Protic ionic liquids (PILs) have been widely employed with the label of "green solvents'' in different sectors of technology and industry. The studied PILs are promising for corrosion inhibition and lubrication applications in industry. Industrial use of the PILs can transform them in wastes, due to accidental spill or drag in water due to washing, that can reach water bodies. In addition, the handling of the product by the workers can expose them to accidental contact. Thus, the aim of this work is to evaluate the toxicity of PILs 2-hydroxyethylammonium oleate (2-HEAOl), N-methyl-2-hydroxyethylammonium oleate (m-2HEAOl) and bis-2-hydroxyethylammonium oleate (BHEAOl) towards Escherichia coli, zebrafish embryos, model organisms that can be present in water, and human skin cells. This is the first work reporting toxicity results for these PILs, which constitutes its novelty. Results showed that the studied PILs did not inhibit E. coli bacterial growth but could cause human skin cells death at the concentrations of use. LC50 values for zebrafish eggs were 40.21 mg/L for 2HEAOl, 12.92 mg/L for BHEAOl and 32.74 mg/L for m-2HEAOl, with sublethal effects at lower concentrations, such as hatching retarding, low heart rate and absence of free swimming.

The role of stem cell-derived exosomes in the repair of cutaneous and bone tissue.

Exosomes are extracellular vesicles secreted by various cell types, which play important roles in physiological processes. In particular, stem cell-derived exosomes have been shown to play crucial functions in intercellular communication during the tissue healing process. This review summarizes the effects of exosomes derived from different stem cell sources on the repair of cutaneous and bone tissue, focusing on the different pathways that could be involved in the regeneration process. The biogenesis, isolation, and content of exosomes have also been discussed. The effectiveness of exosomes is broadly demonstrated for skin and bone regeneration in animal models, supporting the basis for clinical translation of exosomes as a ready-to-use cell-free therapeutic for skin and bone regeneration.

Dissolution, bioactivity behavior, and cytotoxicity of 19.58Li2 O·11.10ZrO2 ·69.32SiO2 glass-ceramic.

Glass and bioactive glass-ceramic can be used in several applications. In bone growth where good bone/biomaterial adhesion was required, bioactive coatings for implants can improve bone formation. The glass and glass-ceramics of the LZS (Li2 O-ZrO2 -SiO2 ) system are very interesting because of their mechanical, electrical, and thermal properties. Very recently, their biological response in contact with human osteoblast has been evaluated. However, despite several initiatives, there are still no studies that systematically assess this system's bioactivity, dissolution, and cytotoxicity in vitro. This work aims to investigate the dissolution, bioactivity behavior, and cytotoxicity of LZS glass-ceramic. LZS glass-ceramics were produced from SiO2 , Li2 CO3, and ZrSiO4 by melting followed by quenching. The obtained glass frits were milled and uniaxially pressed and heat-treated at 800 and 900°C and submitted to physical-chemical, structural and mechanical characterization. Their dissolution behavior was studied in Tris-HCl, while bioactivity was performed in simulated solution body fluid (SBF). The cytotoxicity test was performed using glass-ceramic in direct contact with mesenchymal stem/stromal cells (SC) isolated from human exfoliated deciduous teeth. Structural and microstructural analyzes confirmed bioactivity. The results show that it was possible to produce bioactive glass-ceramic from LZS, proven by the formation of new calcium phosphate structures such as hydroxyapatite on the surface of the samples after exposure to SBF. The SC viability test performed indicated that the materials were not cytotoxic at 0.25, 0.5, and 1.0 mg/ml. The glass-ceramic system under study is very promising for a medicinal application that requires bioactivity and/or biocompatibility for bone regeneration.

Characterization, Cytotoxicity and Anti-Inflammatory Effect Evaluation of Nanocapsules Containing Nicotine.

(1) Background: Nanotechnology is an emerging field that can be applied in the biomedical area. In this study, Eudragit nanocapsules (NCs) containing nicotine were produced. Nicotine is the main alkaloid found in tobacco and has anti-inflammatory properties. NCs containing nicotine may be used as an adjuvant therapy in the treatment of inflammation in the central nervous system. (2) Methods: Nanocapsules were prepared by the interfacial deposition of the pre-formed polymer method and characterized in terms of zeta potential, diameter, polydispersity index, pH, encapsulation efficiency (EE), stability and sustained release profile. In vitro tests with the PC12 cell line were performed, such as MTT, LIVE/DEAD and ELISA assays, to verify their cytotoxic and anti-inflammatory effects. (3) Results: The nanocapsules presented satisfactory values of the characterization parameters; however, poor encapsulation was obtained for nicotine (8.17% ± 0.47). The in vitro tests showed that the treatment with nanocapsules reduced cell viability, which suggests that the Eudragit or the amount of polymer on top of the cells may be detrimental to them, as the cells were able to survive when treated with bulk nicotine. ELISA showed an increment in the expression of IL-6 and IL-1ß, corroborating the hypothesis that NCs were toxic to the cells because of the increase in the levels of these pro-inflammatory cytokines. (4) Conclusions: This study demonstrates that NCs of Eudragit present toxicity. It is therefore necessary to improve NC formulation to obtain better values for the encapsulation efficiency and reduce toxicity of these nanodevices.

Development of a conduit of PLGA-gelatin aligned nanofibers produced by electrospinning for peripheral nerve regeneration.

A promising alternative to conventional nerve grafting is the use of artificial grafts made from biodegradable and biocompatible materials and support cells. The aim of this study has been to produce a biodegradable nerve conduit and investigate the cytocompatibility with stem cells and its regeneration promoting properties in a rat animal model. A poly (lactic-co-glycolic acid) (PLGA) conduit of aligned nanofibers was produced by the electrospinning method, functionalized with gelatin and seeded either with mouse embryonic stem cells (mESCs) or with human mesenchymal stem cells (SHED). The cell proliferation and viability were analyzed in vitro. The conduits were implanted in a rat model of sciatic nerve lesion by transection. The functional recovery was monitored for 8 weeks using the Sciatic Functional Index (SFI) and histological analyses were used to assess the nerve regeneration. Scaffolds of aligned PLGA fibers with an average diameter of 0.90 ± 0.36 µm and an alignment coefficient of 0.817 ± 0.07 were produced. The treatment with gelatin increased the fiber diameter to 1.05 ± 0.32 µm, reduced the alignment coefficient to 0.655 ± 0.045 and made the scaffold very hydrophilic. The cell viability and Live/dead assay showed that the stem cells remained viable and proliferated after 7 days in culture. Confocal images of phalloidin/DAPI staining showed that the cells adhered and proliferated widely, in fully adaptation with the biomaterial. The SFI values of the group that received the conduit were similar to the values of the control lesioned group. In conclusion, conduits composed of PLGA-gelatin nanofibers were produced and promoted a very good interaction with the stem cells. Although in vitro studies have shown this biomaterial to be a promising biomaterial for the regeneration of nerve tissue, in vivo studies of this graft have not shown significant improvements in nerve regeneration.

Nanotechnology for the Treatment of Spinal Cord Injury.

Spinal cord injury (SCI) affects the central nervous system (CNS) and there is currently no treatment with the potential for rehabilitation. Although several clinical treatments have been developed, they are still at an early stage and have not shown success in repairing the broken fiber, which prevents cellular regeneration and integral restoration of motor and sensory functions. Considering the importance of nanotechnology and tissue engineering for neural tissue injuries, this review focuses on the latest advances in nanotechnology for SCI treatment and tissue repair. The PubMed database was used for the bibliographic survey. Initial research using the following keywords "tissue engineering and spinal cord injury" revealed 970 articles published in the last 10 years. The articles were further analyzed, excluding those not related to SCI or with results that did not pertain to the field of interest, including the reviews. It was observed that a total of 811 original articles used the quoted keywords. When the word "treatment" was added, 662 articles were found and among them, 529 were original ones. Finally, when the keywords "Nanotechnology and spinal cord injury" were used, 102 articles were found, 65 being original articles. A search concerning the biomaterials used for SCI found 700 articles with 589 original articles. A total of 107 articles were included in the discussion of this review and some are used for the theoretical framework. Recent progress in nanotechnology and tissue engineering has shown promise for repairing CNS damage. A variety of in vivo animal testing for SCI has been used with or without cells and some of these in vivo studies have shown successful results. However, there is no translation to humans using nanotechnology for SCI treatment, although there is one ongoing trial that employs a tissue engineering approach, among other technologies. The first human surgical scaffold implantation will elucidate the possibility of this use for further clinical trials. This review concludes that even though tissue engineering and nanotechnology are being investigated as a possibility for SCI treatment, tests with humans are still in the theoretical stage. Impact statement Thousands of people are affected by spinal cord injury (SCI) per year in the world. This type of lesion is one of the most severe conditions that can affect humans and usually causes permanent loss of strength, sensitivity, and motor function below the injury site. This article reviews studies on the PubMed database, assessing the publications on SCI in the study field of tissue engineering, focusing on the use of nanotechnology for the treatment of SCI. The review makes an evaluation of the biomaterials used for the treatment of this condition and the techniques applied for the production of nanostructured biomaterials.

Phloroglucinol derivatives from Hypericum species induce in vitro proliferation of cells involved in the wound healing process.

The genus Hypericum (Hypericaceae) is a recognized source of therapeutic agents, being some species widely used due to their wound healing properties. In a previous study, south Brazilian species H. caprifoliatum, H. carinatum, H. connatum, H. myrianthum and H. polyanthemum demonstrated potential to induce proliferation of keratinocytes. In the present study, the effect of phloroglucinol derivatives isolated from Hypericum on cell proliferation of human keratinocytes, fibroblasts and stem cells was investigated. The best results, determined by the MTT assay, were achieved with cariphenone B at concentrations of 0.01 and 0.1 µM (122.3% and 114%, respectively) on HaCaT cells. Uliginosin B was able to induce the proliferation of mesenchymal stem cells (129% at 10 µM) and MRC5 fibroblasts (152.5% at 5 µM). These findings confirm the capacity of phloroglucinol derivatives to induce the in vitro cellular proliferation and reinforce the importance of Hypericum species as potential sources of wound healing compounds.

HA-hybrid matrix composite coating on Ti-Cp for biomedical application.

Calcium phosphate coatings have been applied to titanium metal substrates and their alloys as a synergistic alternative capable of combining the mechanical properties of metals and the excellent bioactive properties provided by ceramic materials. However, the unsatisfactory adhesion of hydroxyapatite coatings on metallic substrates, as well as their limitation when subjected to mechanical stresses have been reported as a limitation. Biofunctional coatings have been proposed as an alternative to single ceramic coatings, aiming at optimizing the long-term clinical success of biomaterials such as Ti. This work aims at evaluating the morphological properties and biological behavior of Ti-cp coated with matrix composite coating hydroxyapatite-containing hybrid. The hybrid matrix was obtained from TEOS and MTES silicon precursors, with dispersed hydroxyapatite suspended by dip coating. For the morphological characterization FTIR, SEM/FEG, AFM and contact angle measurement were used. Biological behavior was evaluated for toxicity, cell viability and the osteogenic differentiation capacity of mesenchymal stem cells. The composite coatings obtained showed regular dispersion of hydroxyapatite particles in the hybrid matrix, with uniform coating adhering to the Ti-Cp substrate. Nevertheless, although they provided similar viability behavior of mesenchymal stem cells to the Ti-Cp substrate, the evaluated coatings did not present osteoinductive properties. This result is probably due to the pronounced hydrophobic behavior caused by the incorporation of HA.

Development of fibrous PLGA/fibrin scaffolds as a potential skin substitute.

The aim of this study has been to fabricate a hybrid electrospun nanofibrous scaffold composed of poly(lactic-co-glycolic) acid (PLGA)/fibrin polymers to be used as a skin substitute and analyze its physical and biological properties. Fibrin was obtained from rat blood plasma, characterized and solubilized in formic acid. The final electrospinning solution concentration was 40% PLGA (w/v) and 1% fibrin (w/v). To improve spinnability, 3% PEG (w/v) was added. The scaffolds were characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Water contact angle, maximum elongation, thermal stability, degree of swelling, blood compatibility, cytotoxicity and cell viability were analyzed. The characterization by SEM showed randomly oriented nanofibers with a mean diameter of 639.8 ± 241.8 nm for the PLGA/fibrin and 1051.0 ± 290.2 nm for the PLGA. FTIR spectra confirmed the presence of fibrin in the mats. Fibrin incorporation reduced the water contact angle from 118.9 ± 2.9 to 111.1 ± 2.8. The fibrin increased tensile strength and decreased elongation at break. The scaffolds demonstrated blood compatibility and fibrin incorporation improved cell adhesion and viability when direct and indirect MTT analyses were carried out. Thus, it can be concluded that the PLGA/fibrin mat is a promising material for use as a skin substitute.

Vascular Tissue Engineering: Polymers and Methodologies for Small Caliber Vascular Grafts.

Cardiovascular disease is the most common cause of death in the world. In severe cases, replacement or revascularization using vascular grafts are the treatment options. While several synthetic vascular grafts are clinically used with common approval for medium to large-caliber vessels, autologous vascular grafts are the only options clinically approved for small-caliber revascularizations. Autologous grafts have, however, some limitations in quantity and quality, and cause an invasiveness to patients when harvested. Therefore, the development of small-caliber synthetic vascular grafts (<5 mm) has been urged. Since small-caliber synthetic grafts made from the same materials as middle and large-caliber grafts have poor patency rates due to thrombus formation and intimal hyperplasia within the graft, newly innovative methodologies with vascular tissue engineering such as electrospinning, decellularization, lyophilization, and 3D printing, and novel polymers have been developed. This review article represents topics on the methodologies used in the development of scaffold-based vascular grafts and the polymers used in vitro and in vivo.

Galantamine improves functional recovery and reduces lesion size in a rat model of spinal cord injury.

Spinal cord injury (SCI) is a medical condition that currently lacks effective treatment. Galantamine is a reversible, competitive acetylcholinesterase inhibitor, used to treat patients with Alzheimers disease. It has been demonstrated that galantamine increases cerebral neurogenesis and has a neuroprotective effect by binding to nicotinic receptors and has an anti-inflammatory effect due to its allosteric binding to the α7nAChR. In the present study, the effects of galantamine on functional recovery and histological outcome in a rat contusion model of SCI were analyzed. Male Wistar rats were submitted to SCI using a NYU/MASCIS impactor. The animals from the galantamine group were treated with 5 mg/kg galantamine intraperitoneally for 5 days. The Basso, Beattie and Bresnahan scale (BBB) was used to evaluate locomotor activity. The expression of beta3-tubulin, NFM, GFAP, O4, CD68 and CD3 was analyzed by flow cytometry. Rats that received galantamine had significantly higher BBB scores in comparison with the control lesion group. Galantamine treatment increased the percentage of NFM positive cells at 6 weeks post-injury and reduced the size of the lesion. The results indicate that galantamine increased tissue survival and accelerated hind limb motor function recovery. This is the first study that has shown the possibility of therapeutic use of galantamine in a model of acute spinal cord injury.

Repeated three-hour maternal deprivation as a model of early-life stress alters maternal behavior, olfactory learning and neural development.

Early life stress such as physical abuse, trauma or neglect during a critical period of development can elicit negative long-lasting effects on health. Neonatal maternal deprivation (MD) is a stressful event capable of triggering structural and neurobiological changes in Central Nervous System (CNS) development during proliferative and migratory cell differentiation. In this study, we investigated the maternal behavior of lactating rats submitted to protocol of chronic neonatal maternal deprivation (MD) during postnatal day (PND) 1 until 10. We analyzed the effects of the MD in the olfactory memory and cellular proliferation and differentiation in the hippocampus and olfactory bulb in Wistar rat pups on 7, 11 and 21 days postpartum. Analysis in active neurons, cellular differentiation and proliferation, were marked and evaluated by flow cytometry in tissue samples of hippocampi and olfactory bulb. Our results demonstrated an increase in maternal behavior immediately after dam's return to the home-cage in MD group compared to the non-deprived group. In addition, MD pups spent more time (higher latency) to identify the nest odor in comparison to the non-deprived rat pups in the olfactory learning task and showed a significant delay in the neural differentiation and proliferation in the hippocampus and olfactory bulb. These results reveal that disruptions in the mother-infant bonding by the MD induce changes in maternal behavior and interaction with the offspring that could be leading to delayed CNS development and significant impairment in offspring's olfactory learning.

A Report from a Workshop of the International Stem Cell Banking Initiative, Held in Collaboration of Global Alliance for iPSC Therapies and the Harvard Stem Cell Institute, Boston, 2017.

This report summarizes the recent activity of the International Stem Cell Banking Initiative held at Harvard Stem Cell Institute, Boston, MA, USA, on June 18, 2017. In this meeting, we aimed to find consensus on ongoing issues of quality control (QC), safety, and efficacy of human pluripotent stem cell banks and their derivative cell therapy products for the global harmonization. In particular, assays for the QC testing such as pluripotency assays test and general QC testing criteria were intensively discussed. Moreover, the recent activities of global stem cell banking centers and the regulatory bodies were briefly summarized to provide an overview on global developments and issues. Stem Cells 2019;37:1130-1135.