Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM
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Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM. / Müller, I; Kordowich, S; Holzwarth, C; Spano, C; Isensee, G; Staiber, A; Viebahn, S; Gieseke, F; Langer, H; Gawaz, M P; Horwitz, E M; Conte, P; Handgretinger, R; Dominici, M.
in: CYTOTHERAPY, Jahrgang 8, Nr. 5, 01.01.2006, S. 437-44.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Transfer › Begutachtung
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TY - JOUR
T1 - Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM
AU - Müller, I
AU - Kordowich, S
AU - Holzwarth, C
AU - Spano, C
AU - Isensee, G
AU - Staiber, A
AU - Viebahn, S
AU - Gieseke, F
AU - Langer, H
AU - Gawaz, M P
AU - Horwitz, E M
AU - Conte, P
AU - Handgretinger, R
AU - Dominici, M
PY - 2006/1/1
Y1 - 2006/1/1
N2 - BACKGROUND: Multipotent mesenchymal stromal cells (MSC) have become important tools in regenerative and transplantation medicine. Rapidly increasing numbers of patients are receiving in vitro-expanded MSC. Culture conditions typically include FSC because human serum does not fully support growth of human MSC in vitro (MSC(FCS)). Concerns regarding BSE, other infectious complications and host immune reactions have fueled investigation of alternative culture supplements.METHODS: As PDGF has long been identified as a growth factor for MSC, we tested media supplementation with platelet lysate for support of MSC proliferation.RESULTS: We found that primary cultures of BM-derived MSC can be established with animal serum-free media containing fresh frozen plasma and platelets (MSC(FFPP)). Moreover, MSC(FFPP) showed vigorous proliferation that was superior to classical culture conditions containing FCS. MSC(FFPP) morphology was equivalent to MSC(FCS), and MSC(FFPP) expressed CD73, CD90, CD105, CD106, CD146 and HLA-ABC while being negative for CD34, CD45 and surface HLA-DR, as expected. In addition to being phenotypically identical, MSC(FFPP) could efficiently differentiate into adipocytes and osteoblasts. In terms of immune regulatory properties, MSC(FFPP) were indistinguishable from MSC(FCS). Proliferation of PBMC induced by IL-2 in combination with OKT-3 or by PHA was inhibited in the presence of MSC(FFPP).DISCUSSION: Taken together, FCS can be replaced safely by FFPP in cultures of MSC for clinical purposes.
AB - BACKGROUND: Multipotent mesenchymal stromal cells (MSC) have become important tools in regenerative and transplantation medicine. Rapidly increasing numbers of patients are receiving in vitro-expanded MSC. Culture conditions typically include FSC because human serum does not fully support growth of human MSC in vitro (MSC(FCS)). Concerns regarding BSE, other infectious complications and host immune reactions have fueled investigation of alternative culture supplements.METHODS: As PDGF has long been identified as a growth factor for MSC, we tested media supplementation with platelet lysate for support of MSC proliferation.RESULTS: We found that primary cultures of BM-derived MSC can be established with animal serum-free media containing fresh frozen plasma and platelets (MSC(FFPP)). Moreover, MSC(FFPP) showed vigorous proliferation that was superior to classical culture conditions containing FCS. MSC(FFPP) morphology was equivalent to MSC(FCS), and MSC(FFPP) expressed CD73, CD90, CD105, CD106, CD146 and HLA-ABC while being negative for CD34, CD45 and surface HLA-DR, as expected. In addition to being phenotypically identical, MSC(FFPP) could efficiently differentiate into adipocytes and osteoblasts. In terms of immune regulatory properties, MSC(FFPP) were indistinguishable from MSC(FCS). Proliferation of PBMC induced by IL-2 in combination with OKT-3 or by PHA was inhibited in the presence of MSC(FFPP).DISCUSSION: Taken together, FCS can be replaced safely by FFPP in cultures of MSC for clinical purposes.
KW - Antigens, Differentiation
KW - Bone Marrow Cells
KW - Cell Differentiation
KW - Cell Proliferation
KW - Cell Separation
KW - Cells, Cultured
KW - Culture Media, Serum-Free
KW - Humans
KW - Mesoderm
KW - Multipotent Stem Cells
KW - Platelet-Derived Growth Factor
KW - Stromal Cells
U2 - 10.1080/14653240600920782
DO - 10.1080/14653240600920782
M3 - SCORING: Journal article
C2 - 17050248
VL - 8
SP - 437
EP - 444
JO - CYTOTHERAPY
JF - CYTOTHERAPY
SN - 1465-3249
IS - 5
ER -