Improved plaque production for short-term bone marrow and fetal liver cultures.
The addition of 0.5% globulin-free (GF-BSA) or 0.5% delipidated BSA (D-BSA) to short-term murine bone marrow (BM) (cultures) increased the number of plaque-forming cells (PFC) responding to trinitrophenylated lipopolysaccharide (TNP-LPS) 2-5-fold (1.1 X 10(4)-2.7 X 10(4) PFC per 10 X 10(6) nucleated BM cells). Although it was necessary to continue to supplement these cultures with 5% fetal calf serum (FCS), the inclusion of the aforementioned BSA preparations provided enhanced PFC production for all lots of FCS tested. Similarly, these preparations of BSA made it feasible to also culture BM in autologous mouse sera (MS) or in medium without 2-mercaptoethanol (2-ME) if in the latter case the D-BSA was pretreated with 2-ME. Thus, the inclusion of GF-BSA or D-BSA in short term cultures of BM not only substantially increased the number of Ig-secreting B cells produced in response to TNP-LPS but seemed to eliminate the need to screen for supportive batches of FCS or MS. These preparations of BSA also facilitated hapten specific PFC responses of fetal liver cultures.
Medina
C A
CA
Department of Biochemistry, Michigan State University, East Lansing 48824.
Li
G
G
Fraker
P J
PJ
D000818:Animals; D000917:Antibody Formation; D000921:Antibody-Producing Cells / Q000276:immunology*; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D002417:Cattle; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology / Q000196:embryology* / Q000276:immunology; D051379:Mice; D012710:Serum Albumin, Bovine
D003470:Culture Media; D012710:Serum Albumin, Bovine