Quality of stem cell lines for scientific research
ISENET Biobanking understands the importance of assuring quality of stem cell lines for scientific research.
STR profiling analysis performed on the human fetal stem cell line SAI2, p41 using the AuthentiFiler PCR Amplification Kit
Cell Line Authentication/Identity
Cell identity testing is performed on DNA on each cell line and is re-performed after cell line cryopreservation, before distribution.
Human pluripotent stem cell lines are negative for mycoplasma screening by PCR (Venor GeM One Step kit) and Hoechst staining (Mycofluor Mycoplasma Detection Kit)
Sterility and Mycoplasma Testing
ISENET Biobanking performs sterility tests pre/ post thawing to assess the presence of contaminants in the cultures.
These include mycoplasma (PCR analysis, bacterial growth in nutrient media for 3 weeks and indirect DNA staining), bacterial, fungal and viral contamination (NAT technology).
QFQ-banding karyotype analysis of MS-IPSCs (RR16#5, p27) and aCGH analysis performed on SAI2, p41 and H9-hNCPCs, p25
Genetic and Genomic Stability
ISENET Biobanking performs standard karyotyping (QFQ-banding) in order to detect major chromosomal aberrations (greater than 10 Megabases) and translocations. High-throughput array Comparative Genomic Hybridization (aCGH) is carried out to identify copy number variations (CNVs) at genomic level.
Spectral-Karyotyping (SKY)
Spectral-Karyotyping (SKY) is a technique based on the simultaneous visualization of all human chromosomes in different colors. Through state-of-the-art imaging and optical microscope technologies, SKY provides an accurate and simple easy detection of structural and numerical chromosomal aberrations.
Global Methylation Levels of the human Embrionic Stem cell line H9 during striatal differentiation
Epigenetic/Epigenomic Profiling
ISENET Biobanking performs whole genome methylation profiles of cell lines during extended passages and differentiation under appropriate conditions. Epigenomic differences often involve cancer related genes and onco-microRNAs with negative effects on the function, stability, differentiation potential and safe.
Immunofluorescence analysis performed on feta neural stem cells (SAI2, p49)IPSCs (CTR4#9,p29) and HD-IPSCs (ChiPS31, p43) showing pluripotency and/or positivity to differentiation markers
Tripotency, Pluripotency and Differentiation Assays
Immuno-fluorescence in combination with qRT-PCR (data not shown) technologies are used to verify tripotency, pluripotency and differentiation capacity of the stem cell lines under appropriate conditions. Pluripotency and differentiation are tested before and after cryopreservation both at early and late phases.