T/LNSI 011-2024 Active

Main Technical Content: 5. Technical Requirements 5.1 Raw Materials 5.1.1 The raw materials shall comply with the requirements of T/CSCB 0001-2022. 5.1.2 Standards for donor evaluation and screening, collection methods, transportation standards, and handover procedures for donor cells shall be established to ensure the safety of donors and cells. 5.1.3 Donors shall be screened for HIV, HBV, HCV, HTLV, EBV, HCMV, and TP, and the results shall be recorded. 5.2 Preparation and Purification Methods 5.2.1 Human amniotic epithelial stem cells The key quality attributes of human amniotic epithelial stem cells shall comply with the provisions of T/LNSI 002-2023. 5.2.2 Culture supernatant of human amniotic epithelial stem cells To obtain high-quality exosomes, the optimal time for harvesting exosomes is defined as P0-P2 generations of human amniotic epithelial stem cells, requiring a fusion rate of more than 95% in the cell supernatant. 5.2.3 Preparation and purification of exosomes The preparation and purification of exosomes shall comply with the requirements of T/CSCB 0007-2022. 5.3 Key Quality Attributes 5.3.1 Morphology Transmission electron microscopy observation shows that the morphology of exosomes is a cup-shaped structure with a double-membrane structure. 5.3.2 Particle size The particle size of exosomes shall be less than 200 nm, with a peak particle size range of 60-120 nm. 5.3.3 Concentration The concentration of exosomes used for anti-aging of skin and bone joints shall be not less than 3×1010 particles/ml. 5.3.4 Marker proteins Any two of the surface markers CD63, CD81, and CD9 on exosomes shall be positive. 5.3.5 Microorganisms They shall be negative for fungi, bacteria, mycoplasma, HIV, HBV, HCV, HTLV, EBV, HCMV, and TP. 5.3.6 Process control The process control for the preparation of human amniotic epithelial stem cells shall comply with the requirements of T/LNSI 002-2023. The process control for exosome