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LETTER TO THE EDITOR

Cell Culture Medium Composition and Translational Adult Bone Marrow-Derived Stem Cell Research

Imperial College, Faculty of Medicine, Hammersmith Hospital Campus, London, United Kingdom

Correspondence: Ioannis Dimarakis, M.D., Division of Surgery, Oncology, Reproductive Biology and Anaesthetics, Faculty of Medicine, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 ONN, U.K. Telephone: +44-(0)-20-8383-2047; Fax: +44-(0)-20-8383-3212; e-mail: id502{at}imperial.ac.uk

Received on November 22, 2005; accepted for publication on January 18, 2006.

	ABSTRACT

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The growing body of adult bone marrow-derived stem cell research in regenerative medicine has recently entered the clinical transplantation setting. The need for a robust flow of data from basic to translational scientists is imperative. All animal- and/or human-derived products should ideally be excluded and synthetic recombinant alternatives used instead. A variety of reasons make such an approach favorable, including achievement of sounder control levels, reduction of any prion, viral, or zoonose contamination risk, avoidance of possible immunological reactions, as well as decrease in demand on animal supplies. At a preclinical level, investigative cell culture protocols must incorporate serum-free media groups alongside control conditions. Finally, autologous serum should not be neglected until an "off-the-shelf" alternative becomes available in the future.

	INTRODUCTION

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We read with interest the article by Sotiropoulou and colleagues on defining optimal culture conditions for clinical-scale production of human mesenchymal stem cells [1]. Our attention was basically drawn to the fact that although the authors went to the length of assessing eight different culture media, all media were supplemented with fetal calf serum. No groups incorporating serum-free synthetic medium or even human serum (autologous or commercially available) were included in the study design. In a preclinical setting, all animal- and/or human-derived products should ideally be excluded and synthetic recombinant alternatives used instead. Despite the very early recognition and adaptation of this concept in clinical work [2], only recently has wider application been allowed in emerging areas of stem cell research [3, 4]. Mesenchymal stem cell research has not been unscathed by this principle; rat mesenchymal stem cells have been shown to retain their proliferation and differentiation potential when cultured in serum-free medium [5]. Under similar predefined conditions, mesenchymal stem cells were able to undergo directional differentiation toward the neuronal lineage [6].

Multiple reasons make such an approach favorable. Because serum is chemically ill-defined, a reduction of variability in qualitative and quantitative cell culture medium composition secondary to interbatch differences should be expected. This alone will eventually lead to a more robust level of quality control. The risk of prion, viral, or zoonose contamination will also be decreased in parallel with the demand on animals for product supply in accordance with the 3R principle of reduction. Serum contains a variety of proteins that may attach to cells in culture and act as antigenic substrates for immunological reactions once transplanted. This has been shown by Selvaggi et al. when patients infused with lymphocytes cultured in medium supplemented with fetal calf serum developed arthus-like reactions [7]. The detection of newly formed antibodies to fetal calf serum is an indication that immune complex formation followed the cell infusions. In a more relevant clinical study, it has been hypothesized that the contact of skeletal myoblasts with fetal calf serum in culture was the cause of unexplained arrhythmias that led to significant malignant ventricular arrhythmias and sudden deaths in patients [8].

Until a chemically defined serum-free medium is widely available, autologous serum supplementation should not be overlooked. This modality is particularly useful in the clinical setting on a single-patient basis and aids overcoming the above and many ethical considerations. Autologous serum not only has been shown to be indistinguishable from fetal calf serum with regard to both isolation and expansion of human mesenchymal stem cells, but also proved to be superior with respect to osteogenic differentiation [9]. In another study, autologous human serum provided sufficient ex vivo expansion of human bone marrow-derived mesenchymal stem cells while maintaining higher cell motility compared with fetal calf serum [10]. In addition, switching to autologous human serum terminated the side effects associated with immune reactions in the aforementioned studies [7, 8].

It is imperative that as adult bone marrow-derived stem cell research enters the realm of clinical research, guidelines be implemented. There should be no rush to enter the clinical stage if the underlying basic research is not solid; this alone will lead to high-quality translational research. Cell culture medium composition should distance itself from animal products. Although autologous serum is a reliable and safe solution for small clinical studies, it will not be appropriate for future "off-the-shelf" availability of stem cell therapy.

	DISCLOSURES

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The authors indicate no potential conflicts of interest.

	REFERENCES

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1. Sotiropoulou PA, Perez SA, Salagianni M et al. Characterization of the optimal culture conditions for clinical scale production of human mesenchymal stem cells. STEM CELLS 2006;24:462–471.[Abstract/Free Full Text]

2. Holst N, Bertheussen K, Forsdahl F et al. Optimization and simplification of culture conditions in human in vitro fertilization (IVF) and preembryo replacement by serum-free media. J In Vitro Fert Embryo Transf 1990;7:47–53.[Medline]

3. Li Y, Powell S, Brunette E et al. Expansion of human embryonic stem cells in defined serum-free medium devoid of animal-derived products. Biotechnol Bioeng 2005;91:688–698.[CrossRef][Medline]

4. Amit M, Shariki C, Margulets V et al. Feeder layer- and serum-free culture of human embryonic stem cells. Biol Reprod 2004;70:837–845.[Abstract/Free Full Text]

5. Lennon DP, Haynesworth SE, Young RG et al. A chemically defined medium supports in vitro proliferation and maintains the osteochondral potential of rat marrow-derived mesenchymal stem cells. Exp Cell Res 1995;219:211–222.[CrossRef][Medline]

6. Tao H, Rao R, Ma DD. Cytokine-induced stable neuronal differentiation of human bone marrow mesenchymal stem cells in a serum/feeder cell-free condition. Dev Growth Differ 2005;47:423–433.[CrossRef][Medline]

7. Selvaggi TA, Walker RE, Fleisher TA. Development of antibodies to fetal calf serum with arthus-like reactions in human immunodeficiency virus-infected patients given syngeneic lymphocyte infusions. Blood 1997;89:776–779.[Abstract/Free Full Text]

8. Chachques JC, Herreros J, Trainini J et al. Autologous human serum for cell culture avoids the implantation of cardioverter-defibrillators in cellular cardiomyoplasty. Int J Cardiol 2004;95(suppl 1):S29–S33.

9. Stute N, Holtz K, Bubenheim M et al. Autologous serum for isolation and expansion of human mesenchymal stem cells for clinical use. Exp Hematol 2004;32:1212–1225.[CrossRef][Medline]

10. Kobayashi T, Watanabe H, Yanagawa T et al. Motility and growth of human bone-marrow mesenchymal stem cells during ex vivo expansion in autologous serum. J Bone Joint Surg Br 2005;87:1426–1433.

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This Article Abstract Full Text (PDF) All Versions of this Article: 2005-0577v1 24/5/1407    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Dimarakis, I. Articles by Levicar, N. Search for Related Content PubMed PubMed Citation Articles by Dimarakis, I. Articles by Levicar, N.