Cell Fractions from Human Biospecimens
Cureline rsquo;s research team has developed protocols for preparation of primary human viable cells and and snap-frozen cell pellets for in-vitro functional studies and protein and RNA Human RNA , expression analysis
Our scientists can prepare viable primary cells and frozen cell fractions from tumor and normal tissues, blood and bone marrow. If you would like to implement your custom protocol, please contact Cureline rsquo;s Scientific Director, Olga Potapova, Ph.D., to discuss your project specifications.
In order to isolate marker-sepcific human cells, Cureline Inc . operates state-of-the-art fully automated magnetic sorter “autoMACS pro” (Miltenyi Biotec, Germany). This high-throughput tool allows effective separation of the desired fractions of biological substances, including bone marrow, cord blood and peripheral blood. Magnetic sorter and adjacent tools are installed in the secured cell culture facility satisfying BSL-3 criteria. Moreover, ‘extension system’ is employed to relocate sterility-sensitive parts of the sorter to inside of the tissue culture hood (Class II).
Mononuclear cell fractions (such as CD34 , CD133 , CD138 and others) are purified from human bone marrow, cord blood and peripheral blood using manufacturer’s protocol. Cells of interest are frozen as cells pellets in liquid nitrogen (flash-frozen, FF protocol) or as viable cell suspensions in DMSO-containing media (viable cells protocol). All samples are stored in either liquid nitrogen tank or at -80oC freezer.
Automated high-throughput magnetic sorter “autoMACS pro” (Miltenyi Biotec, Germany). Extension tubing system allows moving buffer bottles to inside the tissue culture hood (see right). Cooling system fitting 50-ml, 15-ml or 5-ml tubes is available (behind the glass shield).
Cell material is taken by the mechanical arm of the sorter from the tube placed in the cooling system. A series of up to six programs could be executed in sequence, depending on the type of the tubes used.
Recent Collections
APPLICATION: Functional in vitro studies (signal transduction, apoptosis, cell viability) using a new drug candidate.
Leukemia (CLL, CML, ALL, AML):
Isolation of PBMCs from fresh blood using a Ficoll-based procedure, and preparation of viable human cells in DMSO-containing medium. Collection of matching cancer serum samples and extensive clinical information for each patient.
Chronic Lymphocytic Leukemia (CLL):
Isolation of CD19 human cells cells from fresh blood using magnetic beads and preparation of viable primary cells in DMSO-containing medium. Collection of matching cancer serum samples and extensive clinical information for each patient
Multiple Myeloma (MM):
Preparation of viable primary cultures of CD138 cells isolated from fresh bone marrow aspirate using magnetic beads.
Myelodysplastic Syndrome (MDS):
Preparation of viable human BMMCs, snap-frozen cell pellets and RNA. Collection of matching serum samples and extensive clinical information for each patient
