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作者：Marc Gabriel Berger, Richard Veyrat-Masson, Chantal Rapatel, Stphane Descamps, Jacques Chassagne, Nathalie Boiret-Dupre作者单位：CHU Clermont-Ferrand, GECOM, Hmatologie Biologique, Htel-Dieu, Boulevard Lon Malfreyt, Clermont-Ferrand Cedex , France
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7 Q; O) M0 R" n2 M N9 K6 j" j D 【摘要】6 x9 I/ b f- A, P2 d$ R9 E8 ?/ A
For most therapeutic strategies using MSC, the preliminary amplification is carried out in media containing fetal calf serum (FCS). The theoretical health risk of using a xenogenic serum, a recent practice for which we have limited data, cannot be underestimated, while amplification using human serum (HS) remains controversial. At present, the available information on multipotentiality, self-renewal, and transplantability does not permit the selection of FCS rather than HS. Cellular modifications observed during cell passage seem to indicate a gradual impairment of cells in relation to native MSC, suggesting the making of short cell cultures without necessarily trying to reinfuse a high number of MSC in patients. With this approach, the volume of HS required would remain limited. While clinical studies have already started, many problems remain, such as evaluating the quality of the initial mesenchymal compartment and the biological properties of the cell suspension with FCS compared to those with HS, and depending on culture time. % g' @: }0 E0 c* O9 L0 x
【关键词】 Clinical translation Mesenchymal stem cells Ex vivo expansion Culture8 x/ ~( ?) Q! H2 H
INTRODUCTION
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' j+ E- z2 k/ O; D& zWe have followed with great interest the discussion between P.A. Sotiropoulou's team and Drs. Dimarakis and Levicar suggests that the ex vivo culture phase induces subtle and unexpected modifications in the biology of amplified MSC when the cell product is released and infused into the patient. In theory, using HS, in particular autologous serum, should eliminate or reduce the risk of these secondary effects. Different considerations indicate that we should not rule out the possibility of using HS, especially autologous serum.
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Comparative studies of HS and FCS are not so precise. Most agree in recognizing that from the first passage, autologous HS induces a greater amplification than does FCS .. _7 k1 X$ T! T1 D/ X1 B/ N0 t$ T. h
- _7 U! K; J# \2 ~& p8 b4 m' aCertain factors such as the decrease in multiplication speed, progenitor frequency, telomere length, and multilineage differentiation strongly suggest that cells are impaired when amplified in FCS-containing media, in relation to fresh, non-manipulated cells, and study of these native cells seems essential. One approach may be to consider that the most relevant criterion for choosing a serum should be its ability to obtain MSC with properties as close as possible to those of native MSC. This implies that we have to resolve two problems: (a) identifying progenitors in fresh bone marrow, which involves quality control of the mesenchymal compartment of non-manipulated grafts and (b) evaluating the self-renewal capacity of MSC with suitable tools.$ a1 N! C- G6 a
! s: L! E0 Q$ w/ \6 f m8 T2 q0 @One argument given by P.A. Sotiropoulou et al. for using FCS is the limited amount of available autologous serum. This should be discussed, bearing in mind several factors. (a) The idea that cells need to be amplified before therapeutic use is based on a doubtful rationale: the need to amplify cells because they are rare in bone marrow. Nothing proves that the daughter cells produced by native MSC compensate by their numbers the properties lost or modified during amplification. Conversely, with an allogeneic strategy, amplification remains of great interest for purifying MSC and eliminating the hematopoietic cells involved in the host-graft relation. It is likely that they should be purified by short in vitro amplification, while retaining their properties. Needs for serum would thus be reduced. (b) Culture conditions could be adapted to clinical application. For example, bone reconstruction is a cell therapy, which appears particularly attractive today, from the public health perspective. In this case, the number of required progenitors could be limited and HS would be of particular interest since it seems to favor osteoblast differentiation , and so it seems possible that low serum concentrations, while limiting cell proliferation, may enable production of cells for therapeutic use. This justifies further study., ?# ]7 W+ \: s: J% E! h3 J; }( Y
- M) i! |8 p [* X0 ITo conclude, although clinical studies have already started, many questions remain unsolved. We consider it essential to assess the benefit of HS, autologous and heterologous, for purification and/or amplification of MSC, depending on the clinical application. Relying on the experience we have with hematopoietic stem cells for cell therapy, we must now define a quality control for bone marrow grafts, identify native mesenchymal progenitors, and evaluate the effects of amplification to assess both autologous and allogeneic HS in comparison with FCS to make an informed choice.
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DISCLOSURES+ A0 N/ C( y6 v% X. u3 y
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The authors indicate no potential conflicts of interest.
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发表于 2009-3-4 23:56
发表于 2009-6-6 20:20
