BAF CELL的特性和用途

问：BAF cell的来源是什么？有什么特性和用途呢？

答：ATCC上没有你所谓的BAF cell，DSMZ上有两种BAF cell，如下：

Cell line:BA-F8

Cell type:mouse hybridoma(anti-rat fiber/myosin)

Origin:this hybridoma cell line was described to secrete a mouse monoclonal antibody(IgG2b)that reacts with rat type 1 fibers in immunofluorescence and with rat type 1 MHCs (myosin heavy chain)in Western blotting;the cell line was established in 1986 by fusion of NSO myeloma cells with spleen cells from a BALB/c mouse that had been immunized with purified myosin from bovine atrium

Morphology:single,round cells in suspension(some cells adhere partly,but trypsin is not necessary to detach these cells)

Medium:90% Dulbecco's MEM (4.5 g/L glucose)+10% horse serum+2mM L-glutamine

Subculture:split saturated culture 1:3 to 1:4 every 2-3 days;seed out at ca.0.2x106 cells/ml

Incubation:at 37°C with 5% .

Doubling time:ca.30 hours.

Harvest:maximal density at about 0.8x106 cells/ml.

Storage:frozen with 70% medium,20% FBS,10% DMSO at about 3-5x106 cells/ampoule.

Mycoplasma:negative in DAPI,microbiological culture, RNA hybridization,PCR assays.

Immunology:secretion of a IgG{2}monoclonal antibody.

Species:confirmed as mouse with IEF of AST,NP and by species PCR.

Cytogenetics:murine hypertetraploid/pentaploid karyotype-92(89-99)<4n>-1-3 centric.

fusion markers and 1-3 dmin present in most cells.

Viruses: ELISA: reverse transcriptase positive.

Cell line:BA/F3.

Cell type:mouse pro B cells.

Origin:IL-3 dependent murine pro B cell line established from peripheralblood; apparently derived from BALB/c mouse.

Morphology:mostly single, round(some polymorph)cells in suspension(or occasionally in clumps).

Medium:90% RPMI 1640+10% FBS+10 ng/ml IL-3 or 10%vol conditioned medium of WEHI-3B cell line(DSM ACC 26).

Subculture:split saturated culture 1:10 every 3 days;seed out at about 1-3x105 cells/ml;at high cell density(>2 x 106 cells/ml),a cytokine-independent subclone may grow out relatively quickly.CAVEAT:by favoring selective growth,suboptimal culture of cytokine-dependent cell lines may promote outgrowth of factor-independent subclones.Selective conditions include cytokine insufficiency and cell density.Thus the DSMZ cannot guarantee indefinite stability of factor-dependence of cell lines.

Incubation:at 37°C with 5% CO2.

Doubling time:about 20 hours.

Harvest:maximal density of about 2.5 x 106 cells/ml.

Storage:frozen with 70% medium, 20% FBS, 10% DMSO at about 3 x 106 cells/ampoule.

Mycoplasma:negative in DAPI, microbiological culture, RNA hybridization, PCR assays.

Species:confirmed as mouse with IEF of AST, MDH, NP, PEP B and with Species-PCR.

Cytogenetics:murine near diploid karyotype with 33% polyploidy - 40(36-42)<2n>Molec.

Viruses: ELISA: reverse transcriptase positive.