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Cell biology For more information on these products contact your local VWR sales office, send an e-mail to vwrbiomarke@eu.vwr.com or visit our website www.vwr.com Figure 3: MSCs cultured in BD Mosaic hMSC SF Medium at early and late passages suppress equally to cells grown in traditional 5% serum containing medium. Proliferation measured using Bromodeoxyuridine (BrdU) incorporation and subsequent staining with anti BrdU antibodies and FACS-analysis. Left, population doubling 6, right population doubling 12. Population Doublings (PD) Growth Medium % Suppression 6 BD Mosaic hMSC SF 76 5% Serum 82 12 BD Mosaic hMSC SF 78 5% Serum N/A Extensive benchmarking studies have demonstrated that BD Mosaic hMSC SF Cell Culture Environment shows considerable advantages over current hMSC media by enabling researchers to obtain higher cell numbers in shorter time, while using less media as compared to conventional culture methods. References: 1. Campagnoli C. et al. Identification of mesenchymal stem/ progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood. 98(8):2396. 2001 2. Erices A. et al. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol.109(1):235. 2000 3. Dominici, M. et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8 (4):315. 2006 4. Bartholomew A. et al. Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. Exp Hematol. 30(1):42. 2002 5. Di Nicola M. et al. Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood. 99(10):3838. 2002 Description Pk Cat. No. BD Mosaic system Complete kit: Medium, supplement and surface coating 1 kit 734-2458 Medium 500 ml 734-2459 Growth supplement 8.3 ml 734-2460 Additional products BD Falcon 6 well plates, TC treated, flat bottom with lid 36 734-0054 BD Falcon 175 cm 2 tissue culture flask, TC treated, vented cap 40 734-0047 VWR International I VWRbioMarke Issue 27 I September 2011 I 43
the market source for life science Petakas´ internal environment reflects the O 2 concentration of tissues under normal physiological conditions In vitro culture of cells and tissues requires a controlled environment of both temperature and gas concentrations. In the last 40 years, researchers discovered that exposing cells to variable oxygen conditions affects their physiology in dramatic ways compared to cells kept in traditional hyperoxic (21% of the air) culture conditions. Cells cultured in devices exposed to normal atmospheric concentration of oxygen are traditionally used in basic research, in spite of the artifacts consequent to that hyperoxic environment, however, culturing the cells at lower oxygen concentrations, such as 25 to 40% of the atmospheric concentration (5 to 8% oxygen in air), reduces DNA damage, does not alter cellular size/shape, enhances chromosomal stability and an increases proliferative potential 1 . This reduced oxygen concentration better reflects the O 2 concentration (pO 2 ) of tissues under normal physiological conditions, since most tissues are not exposed to external atmospheric O 2 concentrations (Table 1). Actually, at the cellular level, that excessive O 2 concentration exposure can be significantly toxic 2 . Tissues cells, in mammalians, are exposed to a wide range of O 2 concentrations depending on their blood supply position, capillary flow, red blood cell concentration and hemoglobin content 3 , even within a single organ. For instance, oxygen tensions within the liver vary from about 30 - 40 mm Hg in the liver lobule periphery (portal domain, close to the hepatic artery capillaries) and about 15 mm Hg in the liver lobule centres (suprahepatic domain) 4 . Moreover, It is established that cells in different phases of the cycle consume different quantities of oxygen, for example HeLa cells in S phase consume about 240% more than the same cells in G1, and in mitosis O 2 consumption is slightly lower than in G1 6 . Therefore depending on the growing state of the culture (proportion of cells in S phase) the O 2 needs of a culture are different. What today is still sometimes called mild hypoxia in cell culture (5% O 2 in the incubator atmosphere) only can be identified as absence of typical hyperoxic classic cell cultures. It is known that actual normoxia, to be an inducer of large numbers of genes, not restricted only to those directly involved in the physiological response to hypoxia, such as those required for haematopoiesis and angiogenesis 7,8 . It is becoming progressively clearer that for the in vitro culture of many cell types, consistent exposure to O 2 concentrations well below normal 21% atmospheric O 2 concentration is important. For instance culturing fetal lung fibroblasts 9 , embryonic stem cells 1,10 , chondrocytes 11 , mesenchymal stem cells 12 , and haematopoietic stem cells 13, 14 . Cell cultures developed in classic cell culture devices such as T flasks, Petri dishes and derivatives, provide metabolic O 2 to the cells through the water. Gases from the environment dissolve first in the water, which transfer the dissolved gas to the cells. Those devices all have two common parts: (1) The gas phase in contact with (2) the liquid phase. A large interface guarantees the gas transfer in-between the two. The gas phase, in regular culture applications, is the open atmosphere with air containing up to 21% O 2 . Gas diffusion coefficient, temperature and composition of the media force the formation of an O 2 concentration gradient with a high partial pressure (pO 2 = 140 - 150 mm Hg) on the surface layer and a low concentration layer (pO 2 = 70 - 100 mm Hg) at the deepest level (2 - 4 mm), where the cells are anchored. Therefore, to achieve the needed physiological concentrations in the cell culture media, a Table 1. Petaka is the one and only cell culture device autonomously providing Oxygen concentrations in the media at physiological levels, either for normal tissues or tumor cells. 44 I VWR International I VWRbioMarke Issue 27 I September 2011
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