Services
- Isolation of lymphocytes without Transformation
- Transformation of B-Lymphocytes
- Establishment of fibroblasts from Tissue Biopsies
- Biostorage
- Shipping
- Preparation of seed stocks of cells
- Preparation of cells for experiments
- Cloning of Eukaryotic Cells
- Mycoplasma Testing
Isolation of lymphocytes without Transformation:
Lymphocyte separation, Cell Count, Viability Determination, Cryopreservation of whole blood (1 ml), Cryopreservation of 1-4 vials of non-transformed lymphocytes @ 5 x 106 cells, Database entry for all aliquots.
Transformation of B-Lymphocytes:
Transformation with bulk growth. Lymphocyte separation, Cell Count, Viability Determination, Cryo-preservation of whole blood (1 ml), Cryopreservation of 1-4 vials of non-transformed lymphocytes @ 5 x 106 cells, Transformation of fresh lymphocytes with Epstein-Barr virus, Cryopreservation of 4 vials of (transformed) lymphoblasts @ 5 x 106 cells, Database entry for all aliquots. Allow 8-10 weeks for completion.
Transformation and hold. Lymphocyte separation, Cell Count, Viability Determination, Cryopreservation of whole blood (1 ml), Cryopreservation of 1-4 vials of non-transformed lymphocytes @ 5 x 106cells, Transformation of fresh lymphocytes with Epstein-Barr virus, Cryopreservation of 4 vials of (transformed) lymphoblasts @5 x 106 cells, Database entry for all aliquots. Allow 8-10 weeks for completion.
Transformation and T25 flask. Lymphocyte separation, Cell Count, Viability Determination, Cryopreservation of whole blood (1 ml), Cryopreservation of 1-4 vials of non-transformed lymphocytes @ 5 x 106cells, Transformation of fresh lymphocytes with Epstein-Barr virus, Cryopreservation of 1 vial of (transformed) lymphoblasts @5 x 106 cells, Database entry for all aliquots. Allow 6 weeks for completion.
Subsequent transformation and growth of lymphocytes. Transformation of cryopreserved lymphocytes with Epstein-Barr virus, Bulk growth (1x108 ) of transformed cells, Cryopreservation of 4 vials of (transformed) lymphoblasts @ 5 x106cells, Database entry for all aliquots. Allow 8-10 weeks for completion.
Subsequent transformation and hold. Transformation of cryopreserved lymphocytes with Epstein-Barr virus, Cryopreservation of 4 vials of (transformed) lymphoblasts @ 5 x 106 cells, Database entry for all aliquots. Allow 6-8 weeks for completion.
Subsequent transformation and T25 flask. Transformation of cryopreserved lymphocytes with Epstein-Barr virus, Cryopreservation of 1 vial of (transformed) lymphoblasts @ 5 x 106 cells, Database entry for all aliquots. Allow 6 weeks for completion.
Subsequent growth. Bulk growth (1x108 ) of a transformed cell line, Cryopreservation of cell pellet or 20 vials of transformed lymphoblasts @ 5 x 106 cells, Database entry for all aliquots. Allow 3-4 weeks for completion.
Subsequent growth. T25 flask (5x106 ) of a transformed cell line. Allow 7-10 days for completion.
Establishment of fibroblasts from Tissue Biopsies:
Primary fibroblast cultures will be established from biopsies. Four (4)vials of cells will be cryopreserved (5 x 106 cells per vial) at early passages. This process takes approximately 8-10 weeks.
The Cell Center is a member of the International Society for Biological & Environmental Repositories (ISBER) and follows ISBER's best practices for repositories. Cryostorage capacity includes 170 K and 24 K spaces in liquid nitrogen and -80°C freezers, respectively. Excess capacity (two 40K liquid nitrogen freezers and one -80°C freezer) is available off campus at BioStorage, LLC in Arbutus, MD.
Sample aliquots are stored in at least two separate freezers as an additional safeguard. A relational data base tracks specimens in the freezer inventory system. Freezers are monitored twice daily. All freezers have full maintenance contracts and are connected to a remote alarm system that will contact lab personnel at home during off hours. Storage reports are sent out quarterly. Access is from 10:00-11:30 am & 2:00-4:00 pm. Forty-eight hours’ notice is requested to retrieve 50 or more samples. To retrieve samples that are stored offsite, requests should be placed on Monday or Tuesday; the samples will be available at the Cell Center on Thursday or Friday.
The Cell Center will ship biological specimens. The service includes dry ice, shipping boxes, contact with courier for pick-up and telephone or e-mail follow-up with recipient upon shipping. All Cell Center personnel have been trained in Department of Transportation (DOT) bio-material shipping regulations. Investigators are responsible for preparing material transfer agreements (MTA) through the Johns Hopkins Office of Licensing and Technology. View our flyer here.
Preparation of seed stocks of cells:
Cells (5x106/vial) are cryopreserved in 10% DMSO. All cell lines are tested for mycoplasma prior to freezing.
Preparation of cells for experiments:
Cells can be plated in tissue culture dishes, well plates, T-flasks, spinner flasks or roller bottles. Form 60 records special handling instructions for each to meet the specific assay needs. Cells must be mycoplasma free (see MycoplasmaTesting). Prices vary with flask size.
Eukaryotic cells are cloned using established protocols. Cells must be mycoplasma free (see Mycoplasma Testing). A minimum of 20 clones are generated. Additional clones may be available for an additional fee.
We prepare and ship your cell sample for DNA fluorochrome staining at Bionique Testing Laboratories, Inc. You will receive test results in 48 hours. (See protocol link for samples preparation before submiting them for testing).
This service may not be used for clinical applications. Cells being tested should not be used for human/animal diagnostic or therapeutic use. Potential pathogens should be disclosed to the Cell Center personnel prior to processing.