Kadouri A, Kompier R, Honigwachs-Sha'anani J, Toledo J, Brosh N, Sternberg D, Levy A, Tzehoval E, Zipori D
Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Int J Cell Cloning. 1992 Sep;10(5):299-308. doi: 10.1002/stem.5530100508.
Stromal cells of bone marrow origin produce a variety of known cytokines and some factors exhibiting apparently new biological activities. Several of these were identified by the study of cell to cell interactions and were not found in detectable amounts in media conditioned by the cells. We describe here a culture system that enables the release of stromal cytokines into medium free of any added proteins and supplemented with peptides from casein hydrolysate (0.1%). The absence of serum proteins allows extensive concentration and monitoring of activities that are otherwise undetectable. Stromal cells of the MBA-2.1 clonal cell line were seeded in a stationary bed reactor packed with a carrier of non-woven fabric matrix. After a proliferation phase with serum containing medium, the cells were maintained for over 10 months in protein-free medium. Throughout this extended incubation in the absence of serum or serum replacing proteins, stromal cells retained their viability and continuously released transforming growth factor-beta (TGF-beta), macrophage-colony stimulating factor (M-CSF) and restrictin-P, a cytotoxic factor that specifically arrested the growth of plasmacytoma cells. In addition, interleukin-6 (IL-6) was first undetectable, and later in culture its titer reached a maximum of 180,000 international units (IU)/ml. Concomitantly, the production of restrictin-P diminished and reached its lowest levels at the end of 10 months. The results may imply a possible causal relationship between the expression of IL-6 and restrictin-P, since no similarly significant changes were observed in the titers of M-CSF and TGF-beta. This novel bioreactor system may be adaptable for efficient production of different cytokines under absolute serum-free conditions.
骨髓来源的基质细胞可产生多种已知的细胞因子以及一些具有明显新生物学活性的因子。其中一些因子是通过细胞间相互作用的研究鉴定出来的，在细胞条件培养基中未检测到其存在。我们在此描述一种培养系统，该系统能够使基质细胞因子释放到不含任何添加蛋白且添加了酪蛋白水解物肽（0.1%）的培养基中。血清蛋白的缺失使得能够对原本无法检测到的活性进行大量浓缩和监测。将MBA - 2.1克隆细胞系的基质细胞接种到填充有无纺布基质载体的固定床反应器中。在用含血清培养基进行增殖阶段后，细胞在无蛋白培养基中维持培养超过10个月。在整个无血清或无血清替代蛋白的延长培养过程中，基质细胞保持其活力并持续释放转化生长因子 - β（TGF - β）、巨噬细胞集落刺激因子（M - CSF）和restrictin - P，restrictin - P是一种细胞毒性因子，可特异性地抑制浆细胞瘤细胞的生长。此外，白细胞介素 - 6（IL - 6）最初无法检测到，后来在培养过程中其滴度最高达到180,000国际单位（IU）/ml。同时，restrictin - P的产生减少，并在10个月末达到最低水平。这些结果可能意味着IL - 6和restrictin - P的表达之间可能存在因果关系，因为在M - CSF和TGF - β的滴度中未观察到类似的显著变化。这种新型生物反应器系统可能适用于在绝对无血清条件下高效生产不同的细胞因子。
