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1. Retrieval of germinal zone neural stem cells from the cerebrospinal fluid of premature infants with intraventricular hemorrhage

Author

Fernandez-Munoz, B, Rosell-Valle, C, Ferrari, D, Alba-Amador, J, Montiel, M, Campos-Cuerva, R, Lopez-Navas, L, Munoz-Escalona, M, Martin-Lopez, M, Profico, D, Blanco, M, Giorgetti, A, Gonzalez-Munoz, E, Marquez-Rivas, J, Sanchez-Pernaute, R, Fernandez-Munoz B., Rosell-Valle C., Ferrari D., Alba-Amador J., Montiel M. A., Campos-Cuerva R., Lopez-Navas L., Munoz-Escalona M., Martin-Lopez M., Profico D. C., Blanco M. F., Giorgetti A., Gonzalez-Munoz E., Marquez-Rivas J., Sanchez-Pernaute R., Fernandez-Munoz, B, Rosell-Valle, C, Ferrari, D, Alba-Amador, J, Montiel, M, Campos-Cuerva, R, Lopez-Navas, L, Munoz-Escalona, M, Martin-Lopez, M, Profico, D, Blanco, M, Giorgetti, A, Gonzalez-Munoz, E, Marquez-Rivas, J, Sanchez-Pernaute, R, Fernandez-Munoz B., Rosell-Valle C., Ferrari D., Alba-Amador J., Montiel M. A., Campos-Cuerva R., Lopez-Navas L., Munoz-Escalona M., Martin-Lopez M., Profico D. C., Blanco M. F., Giorgetti A., Gonzalez-Munoz E., Marquez-Rivas J., and Sanchez-Pernaute R.

Abstract

Intraventricular hemorrhage is a common cause of morbidity and mortality in premature infants. The rupture of the germinal zone into the ventricles entails loss of neural stem cells and disturbs the normal cytoarchitecture of the region, compromising late neurogliogenesis. Here we demonstrate that neural stem cells can be easily and robustly isolated from the hemorrhagic cerebrospinal fluid obtained during therapeutic neuroendoscopic lavage in preterm infants with severe intraventricular hemorrhage. Our analyses demonstrate that these neural stem cells, although similar to human fetal cell lines, display distinctive hallmarks related to their regional and developmental origin in the germinal zone of the ventral forebrain, the ganglionic eminences that give rise to interneurons and oligodendrocytes. These cells can be expanded, cryopreserved, and differentiated in vitro and in vivo in the brain of nude mice and show no sign of tumoral transformation 6 months after transplantation. This novel class of neural stem cells poses no ethical concerns, as the fluid is usually discarded, and could be useful for the development of an autologous therapy for preterm infants, aiming to restore late neurogliogenesis and attenuate neurocognitive deficits. Furthermore, these cells represent a valuable tool for the study of the final stages of human brain development and germinal zone biology.

Published
2020

2. Retrieval of germinal zone neural stem cells from the cerebrospinal fluid of premature infants with intraventricular hemorrhage

Author

Manuel Francisco Blanco, Daniela Celeste Profico, Elena González-Muñoz, Beatriz Fernández-Muñoz, Miguel Ángel Montiel, Cristina Rosell-Valle, María Muñoz-Escalona, Rosario Sanchez-Pernaute, María Martín-López, Julia Alba-Amador, Rafael Campos-Cuerva, Javier Márquez-Rivas, Daniela Ferrari, Alessandra Giorgetti, Luis Lopez‐Navas, Fernandez-Munoz, B, Rosell-Valle, C, Ferrari, D, Alba-Amador, J, Montiel, M, Campos-Cuerva, R, Lopez-Navas, L, Munoz-Escalona, M, Martin-Lopez, M, Profico, D, Blanco, M, Giorgetti, A, Gonzalez-Munoz, E, Marquez-Rivas, J, Sanchez-Pernaute, R, Junta de Andalucía, Ministerio de Economía y Competitividad (España), Generalitat de Catalunya, [Fernández-Muñoz,B, Rosell-Valle,C, Alba-Amador,J, Montiel,MÁ, Campos-Cuerva,R, Muñoz-Escalona,M, Martín-López,M, Blanco,MF] Unidad de Producción y Reprogramación Celular (UPRC), Red Andaluza para el diseño y traslación de Terapias Avanzadas, Sevilla, Spain. [Fernández-Muñoz,B, Márquez-Rivas,J] Grupo de Neurociencia aplicada, Instituto de Biomedicina de Sevilla, Sevilla, Spain. [Ferrari,D] Department of Biotechnology and Biosciences, University Milan-Bicocca, Milan, Italy. [Campos-Cuerva,R] Centro de Transfusiones, Tejidos y Células de Sevilla (CTTS), Sevilla, Spain. [Lopez-Navas,L, Sanchez-Pernaute,R] Departamento de Preclínica, Red Andaluza de Diseño y Traslación de Terapias Avanzadas, Sevilla, Spain. [Profico,DC] Fondazione IRCCS Casa Sollievo della Sofferenza, Production Unit of Advanced Therapies (UPTA), San Giovanni Rotondo, Italy. [Giorgetti,A] Regenerative Medicine Program, Bellvitge Biomedical Research Institute (IDIBELL), Program for Translation of Regenerative Medicine in Catalonia (P-CMRC), Barcelona, Spain. [González-Muñoz,E] Department of Cell Biology, Genetics and Physiology, University of Málaga, Málaga, Spain. [González-Muñoz,E] Department of Regenerative Nanomedicine, Andalusian Center for Nanomedicine and Biotechnology-BIONAND, Málaga, Spain. [González-Muñoz,E] Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN). Carlos III Health Institute (ISCIII), Spain. [Márquez-Rivas,J] Neurosurgery Department, Hospital Virgen del Rocío, Sevilla, Spain., and This work was supported by research funds from the Andalusian Consejería de Salud to the Red Andaluza de Diseño y Traslación de Terapias Avanzadas with contribution from the COST Action CA16122 for STSM and networking. AG is supported by Ramon y Cajal Program (RyC-2013-13221), MINECO(SAF2016-80205-R) and CERCA Pro gram/Generalitat de Catalunya.

Subjects
0301 basic medicine, Male, Neurobiologia del desenvolupament, Pathology, Premature infant, Infants prematurs, neural stem cell, 0302 clinical medicine, Cerebrospinal fluid, Neural Stem Cells, Tissue‐specific Progenitor and Stem Cells, Organisms::Eukaryota::Animals [Medical Subject Headings], AC133 Antigen, Developmental neurobiology, Hemorragia cerebral intraventricular, Anatomy::Cells::Stem Cells::Neural Stem Cells [Medical Subject Headings], lcsh:R5-920, lcsh:Cytology, Premature infants, Líquido cefalorraquídeo, Neurogenesis, Gene Expression Regulation, Developmental, General Medicine, Human brain, Recién nacido prematuro, Neural stem cell, premature infant, Anatomy::Fluids and Secretions::Body Fluids::Extracellular Fluid::Cerebrospinal Fluid [Medical Subject Headings], Persons::Persons::Age Groups::Infant::Infant, Newborn::Infant, Premature [Medical Subject Headings], Intraventricular hemorrhage, medicine.anatomical_structure, Centro germinal, Female, lcsh:Medicine (General), Infant, Premature, medicine.medical_specialty, neurogenesi, Mice, Nude, Check Tags::Male [Medical Subject Headings], Phenomena and Processes::Genetic Phenomena::Genetic Processes::Gene Expression Regulation::Gene Expression Regulation, Developmental [Medical Subject Headings], intraventricular hemorrhage, cerebrospinal fluid, 03 medical and health sciences, medicine, Animals, lcsh:QH573-671, Cerebral Hemorrhage, Células madre nerviosas, business.industry, Germinal zone, germinal zone, Diseases::Nervous System Diseases::Central Nervous System Diseases::Brain Diseases::Cerebrovascular Disorders::Intracranial Hemorrhages::Cerebral Hemorrhage [Medical Subject Headings], BIO/13 - BIOLOGIA APPLICATA, Líquid cefalorraquidi, Endoscopy, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Mice::Mice, Mutant Strains::Mice, Nude [Medical Subject Headings], Cell Biology, medicine.disease, Transplantation, 030104 developmental biology, Check Tags::Female [Medical Subject Headings], Cell culture, Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Diagnostic Techniques, Surgical::Endoscopy [Medical Subject Headings], Forebrain, business, 030217 neurology & neurosurgery, Developmental Biology
Abstract

Intraventricular hemorrhage is a common cause of morbidity and mortality in premature infants. The rupture of the germinal zone into the ventricles entails loss of neural stem cells and disturbs the normal cytoarchitecture of the region, compromising late neurogliogenesis. Here we demonstrate that neural stem cells can be easily and robustly isolated from the hemorrhagic cerebrospinal fluid obtained during therapeutic neuroendoscopic lavage in preterm infants with severe intraventricular hemorrhage. Our analyses demonstrate that these neural stem cells, although similar to human fetal cell lines, display distinctive hallmarks related to their regional and developmental origin in the germinal zone of the ventral forebrain, the ganglionic eminences that give rise to interneurons and oligodendrocytes. These cells can be expanded, cryopreserved, and differentiated in vitro and in vivo in the brain of nude mice and show no sign of tumoral transformation 6 months after transplantation. This novel class of neural stem cells poses no ethical concerns, as the fluid is usually discarded, and could be useful for the development of an autologous therapy for preterm infants, aiming to restore late neurogliogenesis and attenuate neurocognitive deficits. Furthermore, these cells represent a valuable tool for the study of the final stages of human brain development and germinal zone biology., Germinal zone neural stem cells (Gz‐NSC) are isolated from the hemorrhagic cerebrospinal fluid of preterm infants with severe intraventricular hemorrhage. These cells express ventral and posterior forebrain markers, can be differentiated and do not cause tumors. Gz‐NSC represent a valuable tool for the development of new cell therapies and the study of human Gz biology.

Published
2020

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