Growth of myeloma cell lines

Grow mouse myelomas in static flasks or spinner culture and keep growing exponentially in DMEM containing 10% or 20% FCS (dilute to 2-3 x 10 ceUs/ml the day before fusion). 

The rat myeloma Y3 has to be grown in spinner culture to fuse weU. Seven to ten days before cells are required, about 5 x 10 cells stored frozen in liquid nitrogen as 1 ml aliquots in freezer medium are thawed quickly at 37 °C, diluted with 10 ml 

The Y3 myeloma has a generation time of about 10-12 h and exponentially grovwng cultures require feeding daily by fomfold dilution writh fresh medium. 

Preparation of Immune lymphocytes and myeloma cells for fusion 

Mice or rats taken three days after final immimization 

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Billadeau, D., Liu, P., Jelinek, D., Shah, N., LeBien, T. W. and van Ness, B. (1997). Activating mutations in the N- and K-ras oncogenes differentially affect the growth properties of the IL-6-dependent myeloma cell line ANBL6. Cancer Res. 57, 2268-2275. 

Several s mtheses of nucleotidephospholipids have been reported recently. For example, phospholipid-araC conjugates have been prepared and tested as prodrugs of arotC as inhibitors of the growth of a murine myeloma cell line. The most effective inhibitor was [13, = 1, R = Me(CH2)i4] solubility difficulties may have affected the testing of other analogues. 

Passive antibody vaccines have been prepared up to now from human blood serum. Consequently, there has been no need for cultivation methods beyond vaccination and conventional harvest of antibody-containing blood from donors. Due to safety concerns over using human blood, passive vaccines will likely be monoclonal antibodies or cocktails thereof prepared in vitro by the cultivation of hybridoma or myeloma cell lines. This approach is under investigation for anti-HIV-1 antibodies [Emini et al., 1992]. Cultivation of these cell lines involves the same principles of animal cell cultivation as described above, with the exception that hybridomas can be less fastidious in nutritional requirements, and they do not require surface attachment for growth. These features will allow for defined serum-free media and simpler cultivation vessels and procedures. 

Another important aspect involved in the selection of transfected lines is the capacity to grow without physical support, since the scale-up of such processes is much simpler than those designed for growth of anchorage-dependent cells. Thus, cells that grow naturally in suspension are preferred, such as myeloma cells (Sp2/0 and NSO), or others that can be easily adapted to this form of cultivation, such as CHO and BHK (Chu and Robinson, 2001). 

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