The NSCs nasal patch complex preparation
hepatitis B, abortion, syphilis, human immunodeficiency virus, chlamydia
HEPATITIS B, HUMAN IMMUNODEFICIENCY VIRUS, SYPHILIS, STERILE, MYCOPLASMA, SEPARATION, COLD
The allogeneic NSCs were isolated from aborted human fetal forebrain tissue. These tissues were collected from healthy pregnant women who had requested an artificial abortion. All the donors voluntarily signed informed consent forms, and the tests for human immunodeficiency virus, syphilis, hepatitis B, and other pathogenic microorganisms were negative. All donor tissue was stored in sealed containers of sterile 0.9% normal saline (NS) and transported to the stem cell preparation center registered with the China Food and Drug Administration (CFDA) via a cold chain within 6 h of abortion. The whole procedure was in accordance with the current international Good Manufacturing Practices process for separation, extraction, amplification, passage, and detection. The NSCs were passaged every 7–10 days and carried out using passages 4–8 for our clinical research. Before clinical application, cytopathic tests were performed for endotoxins, mycoplasma, chlamydia, and viruses. And the proportions of positive antigens expression were calculated by Flow Cytometry assay of NSCs biomarkers (Vimentin, Nestin, Notch-1, SOX2, SSEA1and Musashi-1). Immunofluorescence staining was used to detect the differentiation ability of NSCs into neurons, astrocytes, and oligodendrocytes. Observation of interaction between patches and NSCs was performed by overlaying images of cells and materials and then analyzed by confocal photographing. 2 × 10