Serum-free media formulations are cell line–specific and require optimization for microcarrier culture

The production of human endothelial cell secretory products in amounts sufficient for biochemical studies is largely restricted by the culture growth area. Conventional flat bed systems yield at best 20-30 x 106 cells per 180cm2 culture flask. To overcome this problem, cells may be grown on Cytodex 3 microcarriers allowing large numbers of cells to be grown and conditioned in small culture volumes. A typical microcarrier unit will contain 200-300 x 106 cells and may be expanded in excess of 1000 x 106 cells at confluence. High viability (95%) and recovery (70-80%) in sub-culturing of microcarrier to microcarrier culture can be achieved with careful management of culture conditions and brief exposure to enzymes.Human umbilical artery and vein, and saphenous vein endothelial cells were prepared and grogn on microcarrier cultures to cell populations of 200-450 x 106 cells and conditioned for 14 day periods in serum-free media.The production profiles of several endothelial cell proteins including thrombospondin (TSP), von Willebrand Factor (vWF) and issue plasminogen activator (t-PA) were measured by radioimmunoassay under these conditions, and demonstrate the use of microcarrier cultures in producing milligram quantities of engothelial cell protein. For example, a HUVEC culture of 200 x 106 cells conditioned with serum-free media for 14 days yielded a total of 6.9mg TSP, 0.7mg vWF and 48.9ug t-PA. In this laboratory one such application of the system was the purification of endothelial proteins in amounts sufficient for immunisation of mice prior to the production of monoclonal antibodies and for subsequent characterisation.

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Differentiation of Myoblasts in Serum-Free Media: Effects of Modified Media Are Cell Line-Specific

Cells Tissues Organs ◽

10.1159/000016776 ◽

2000 ◽

Vol 167 (2-3) ◽

pp. 130-137 ◽

Cited By ~ 91

Author(s):

Moira A. Lawson ◽

Peter P. Purslow

Keyword(s):

Cell Line ◽

Media Effects ◽

Serum Free ◽

Free Media ◽

Modified Media

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Serum-free media for a human lymphocyte cell line and for PWM-stimulated peripheral blood lymphocytes: Requirements for insulin, transferrin and albumin

Immunology Letters ◽

10.1016/0165-2478(83)90100-1 ◽

1983 ◽

Vol 6 (3) ◽

pp. 169-174 ◽

Cited By ~ 5

Author(s):

Andrew Yen ◽

Rose Duigou

Keyword(s):

Cell Line ◽

Human Lymphocyte ◽

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Serum Free ◽

Blood Lymphocytes ◽

Free Media ◽

Lymphocyte Cell ◽

Lymphocyte Cell Line

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Isolation, Culture and Identification of Choriocarcinoma Stem-Like Cells from the Human Choriocarcinoma Cell-Line JEG-3

Cellular Physiology and Biochemistry ◽

10.1159/000447845 ◽

2016 ◽

Vol 39 (4) ◽

pp. 1421-1432 ◽

Cited By ~ 12

Author(s):

Jingting Cai ◽

Tianfang Peng ◽

Jing Wang ◽

Jingli Zhang ◽

Hui Hu ◽

...

Keyword(s):

Cell Line ◽

Colony Formation ◽

Soft Agar ◽

Serum Free ◽

Significant Difference ◽

Free Media ◽

Proliferative Ability ◽

Choriocarcinoma Cell Line

Background/Aims: Cancer stem cells (CSCs) exhibit enhanced proliferative capacity and resistance to chemotherapy; however, choriocarcinoma CSCs have not yet been reported. In this study the human choriocarcinoma cell line JEG-3 was cultured in serum free media, and the characteristics of suspension and parental adherent JEG-3 cells were compared. Methods: Cell proliferation, colony-formation, soft agar clonogenicity, and transwell invasion assays were performed in vitro, and tumor xenografts in BALB/c nude mice were used to evaluate stem cell properties. Results: In serum-supplemented medium (SSM), JEG-3 cells were 4.51 ± 1.71% CD44+, 7.67 ± 2.67% CD133+, and 13.85 ± 2.95% ABCG2+. In serum-free medium (SFM), the expression of these markers increased to 53.08 ± 3.15%, 47.40 ± 2.67%, and 78.70 ± 7.16%, respectively. Moreover, suspension JEG-3 cells exhibited enhanced colony-formation capability as well as invasive and proliferative ability in vitro, alongside enhanced tumorigenic properties in vivo. Suspension JEG-3 cells also exhibited resistance to the chemotherapeutic drugs methotrexate, fluorouracil and etoposide. When seeded in serum supplemented medium, suspension JEG-3 cells readopted an adherent phenotype and continued to differentiate with no significant difference in the morphology between suspension and parent cells. Conclusion: In this study, choriocarcinoma stem-like cells (CSLCs) were isolated from the human choriocarcinoma JEG-3 cell line by SFM culture and characterized.

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