Comparison of proteomic profiles of serum, plasma, and modified media supplements used for cell culture and expansion

Saleh Ayache,David F Stroncek,Susan F Leitman,Francesco M Marincola,Stefanie Slezak,Karen M Byrne,Monica C Panelli

doi:10.1186/1479-5876-4-40

Saleh Ayache, David F Stroncek + Show 5 more

Open Access

https://doi.org/10.1186/1479-5876-4-40

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Abstract

BackgroundThe culture and expansion of human cells for clinical use requires the presence of human serum or plasma in culture media. Although these supplements have been extensively characterized in their chemical composition, only recently it has been possible to provide by high throughput protein analysis, a comprehensive profile of the soluble factors contributing to cell survival. This study analyzed and compared the presence of 100 proteins including chemokines, cytokines and soluble factors in six different types of media supplements: serum, plasma, recalcified plasma, heat inactivated serum, heat inactivated plasma and heat inactivated recalcified plasma.MethodsSerum, plasma, recalcified plasma, and heat inactivated supplements were prepared from ten healthy subjects. The levels of 100 soluble factors were measured in each sample using a multiplexed ELISA assay and compared by Eisen hierarchical clustering analysis.ResultsA comparison of serum and plasma levels of soluble factors found that 2 were greater in plasma but 18 factors were greater in serum including 11 chemokines. The levels of only four factors differed between recalcified plasma and plasma. Heat inactivation had the greatest effect on soluble factors. Supervised Eisen hierarchical clustering indicated that the differences between heat inactivated supplements and those that were not were greater than the differences within these two groups. The levels of 36 factors differed between heat inactivated plasma and plasma. Thirty one of these factors had a lower concentration in heat inactivated plasma including 12 chemokines, 4 growth factors, 4 matrix metalloproteases, and 3 adhesion molecules. Heat inactivated decalcified plasma is often used in place of heat inactivated serum and the levels of 19 soluble factors differed between these two supplements.ConclusionOur report provides a comprehensive protein profile of serum, plasma recalcified plasma, and heat inactivated supplements. This profile represents a qualitative and quantitative database that can aid in the selection of the appropriate blood derived supplement for human cell cultures with special requirements.

Highlights

The culture and expansion of human cells for clinical use requires the presence of human serum or plasma in culture media
The use of bovine serum exposes patients to the potential risk of transmission of pathogens such as Creutzfeldt-Jakob disease (CJD) [11] and the development of a humoral or cellular response to bovine proteins [4]. In spite of these issues, some clinical protocols use fetal calf serum [3,6], but often human serum is used in place of fetal calf serum [1,7,8]
The goal of this study was to determine if differences in the levels of soluble factors among these media supplements might contribute to the differences in their efficacy

Summary

Introduction

The culture and expansion of human cells for clinical use requires the presence of human serum or plasma in culture media. The use of bovine serum exposes patients to the potential risk of transmission of pathogens such as Creutzfeldt-Jakob disease (CJD) [11] and the development of a humoral or cellular response to bovine proteins [4]. In spite of these issues, some clinical protocols use fetal calf serum [3,6], but often human serum is used in place of fetal calf serum [1,7,8]. In some cases human plasma [1] or no media supplement is used [2,4,9,10]

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