scholarly journals Human Cells Grown With or Without Substitutes for Fetal Bovine Serum

Cell Medicine ◽  
2018 ◽  
Vol 10 ◽  
pp. 215517901875514 ◽  
Author(s):  
John E. Piletz ◽  
Jennifer Drivon ◽  
John Eisenga ◽  
Will Buck ◽  
Sabrina Yen ◽  
...  

Safety concerns over cell-derived pharmaceutical products being manufactured in supplements of fetal bovine serum (FBS) have ignited pleas to replace FBS. Herein, four newly marketed alternatives to FBS were compared: a xeno-free product called Cell-Ess®, a human platelet lysate marketed as GroPro®, and two mixtures of adult bovine serum varying in their proportions of neonatal growth factors, called Liporo® and FetalGro®. An endothelial cell line (C2BBe1) and a neuronal cell line (SHSY5Y) near confluency in media with 10% FBS were selectively scraped and taken through a 25-day step-wise algorithm to replace FBS, and another human endothelial cell line (HRA-19) was studied to replicate C2BBe1. Cells were stained, counted, and compared for viability, migration, and spheroids. The C2BBe1 and HRA-19 cell lines failed to proliferate in 10% Cell-Ess® but grew in 10% GroPro® or 10% FetalGro® reasonably well compared to reference 10% FBS. With SH-SY5Y, only FetalGro® approached FBS's efficacy. These were all inferior to 11 different branded lots of FBS (positive controls), but five days into switching just amongst the FBS brands, 4 of 11 supported less proliferation than reference FBS in endothelial HRA-19 ( p < 0.004). Moreover, neurospheres were enriched in two branded lots of FBS and FetalGro® (each p < 0.004), neurospheres being an unwanted phenotype for any neuronal cell application. Because platelet-derived GroPro® stood out amongst the non-FBS growth supplements to allow proliferation without inducing spheroids, it seems the best (mindful that the cells still grew slower in it compared to FBS). While no perfect replacement was found amongst the alternatives to FBS, the algorithm for switching should be useful in future testing of new alternatives to FBS as the need arises to switch from FBS and expand pharmaceutical products with safety for human use.

Biomaterials ◽  
2016 ◽  
Vol 76 ◽  
pp. 371-387 ◽  
Author(s):  
Thierry Burnouf ◽  
Dirk Strunk ◽  
Mickey B.C. Koh ◽  
Katharina Schallmoser

2021 ◽  
Author(s):  
Zhuo Zhen Chen

Blood proteins communicate with many different cells, tissue and organs; perform key functions in the immune system and may be of particular biological complexity. One of the most widely used blood products in the laboratory is fetal bovine serum for cell culture. There are ethical and practical concerns regarding the use of fetal serum from animals and alternative serum-free replacements have been attempted using platelet lysates. Previous biochemical experiments have shown that FBS apparently contained factors such as alpha-feto protein (AFP) and insulin-like growth factors that may support the indefinite cell growth and division of certain cell lines. It is presumed that a set of as yet undefined growth factors transform cells growth resulting in rapid proliferation. Cultured Raw cells 264.7 in adult bovine serum multiplied slowly and differentiated into elongated cells with a dendritic shape, which died after the first few generations. On the contrary, in fetal bovine serum, cultured cells multiplied rapidly and formed many smaller cells with a rounded shape through many cell passages. Three independent batches of fetal bovine serum were tested on Raw cells 264.7 macrophages to confirm that they supported cell growth in culture compared to three independent batches of adult bovine serum. The intact proteins of each serum sample were separated by partition chromatography into 16 fractions with an increasing step gradient of salts over quaternary amine resin (proteomics). The endogenous peptides were precipitated with 90% of acetonitrile and extracted into 10 fractions with a decreasing step gradient of acetonitrile in water (peptidomics). Trypsin digested intact proteins and endogenous peptides were then analyzed on a fresh C18 nano-HPLC column with random and independent sampling by LC-ESI-MS/MS. The fractionated mass spectra were identified with SEQUEST and X!TANDEM algorithms. Redundant use of MS/MS spectra were flirted out with the SQL Server system and the R statistical analysis system was used to perform Chi Square (X2) analysis of frequency counts and ANOVA of the log10 precursor intensity results. Alpha-feto protein, fetal albumin, insulin, insulin like growth factors, platelet derived growth factors and proteins associated with HRAS/AKT growth pathway at the level of ligand, receptors, receptor associated enzyme and nucleic acid binding proteins including transcription factors were observed to be specifically enriched in fetal serum.


1969 ◽  
Vol 15 (10) ◽  
pp. 1197-1200 ◽  
Author(s):  
S. S. Sohi

Aedes aegypti cells (Grace's line) growing in a medium containing 5% Antheraea pernyi hemolymph (APH) and 5% fetal bovine serum (FBS) were adapted to a hemolymph-free medium. The quantity of APH was gradually substituted with an equal quantity of FBS over a 2-month period. The cells grew very slowly in the hemolymph-free medium in the beginning. However, they became adapted to it in another 8 months; after that they grew as well as the cells in medium supplemented with hemolymph.Also, these cells grew as well in medium containing Bombyx mori hemolymph as in medium enriched with APH.


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