Universidade Federal de São Paulo (UNIFESP)

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Concomitant Isolation of Primary Astrocytes and Microglia for Protozoa Parasite Infection

Aline de Oliveira Lima Pacheco^*1, Marcelo Pires Amaral^*1, Ingrid Sancho de Farias¹, Luiza Zainotti Miguel Fahur Bottino¹, Karina Ramalho Bortoluci²

¹Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP), ²Departamento de Ciências Biológicas, Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP)

The overall goal of this protocol is to instruct how to extract, maintain, and dissociate murine astrocyte and microglia cells from the central nervous system, followed by infection with protozoa parasites.

Use of LeishGEdit CRISPR-Cas9 Technology for Genetic Manipulation of Protozoan Parasite Leishmania

Suellen Rodrigues Maran¹, Bruno Souza Bonifácio¹, Myrna Zanchetta¹, Miguel Antonio do Nascimento Garcia¹, Carolina M. C. Catta-Preta², Nilmar Silvio Moretti¹

¹Laboratory of Molecular Biology of Pathogens - Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo (UNIFESP), ²Center of Medicinal Chemistry (CQMED), University of Campinas (Unicamp)

The establishment of a robust CRISPR-Cas9 methodology for genetic manipulation of Leishmania has accelerated the understanding of key biological processes of this parasite. Here, we describe in detail all the steps to generate knockout or in situ fluorescent-tagged parasites of virtually any gene of interest using LeishGEdit methodology.

Medicine

Technique for Obtaining Mesenchymal Stem Cell from Adipose Tissue and Stromal Vascular Fraction Characterization in Long-Term Cryopreservation

Leniza Pola-Silva^1,2, Fabio Xerfan Nahas³, Flavia Nascimento², Tatiana Rabelo Santos², Andrea Moraes Malinverni¹, Adelson Alves², Lydia Masako Ferreira³, Maria Isabel Melaragno¹

¹Genetics Division, Universidade Federal de São Paulo (UNIFESP), ²TechLife Biotech, São Lucas Cell Therapy Group, ³Division of Plastic Surgery, Universidade Federal de São Paulo (UNIFESP)

The present protocol describes an improved methodology for ADSC isolation resulting in a tremendous cellular yield with time gain compared to the literature. This study also provides a straightforward method for obtaining a relatively large number of viable cells after long-term cryopreservation.