关于如何区别MSCs和fibroblasts的参考文献

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上海过客的帖子“MEF 到底是一种什么性质的细胞？是单一的细胞？肌纤维细胞，神经纤维细胞？”引起大家的热议，同时也引发一个新的讨论，如何区别MSCs和fibroblasts。我大概检索了一下相关的参考文献，供大家讨论，欢迎补充！% Q% |/ g0 f4 l9 e2 ], G0 X4 U$ [
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1.J Periodontal Res. 2007 Jun;42(3):283-6.% n1 X( ?; L# u- o+ b. e: ?' D

  A* w6 d. V  T6 ZIdentification of marker genes distinguishing human periodontal ligament cells from human mesenchymal stem cells and human gingival fibroblasts.
* p3 a* l3 t2 G- M, hFujita T, Iwata T, Shiba H, Igarashi A, Hirata R, Takeda K, Mizuno N, Tsuji K, Kawaguchi H, Kato Y, Kurihara H.' D* e4 }1 K7 |: n6 L) [
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Department of Periodontal Medicine, Division of Frontier Medical Science, Hiroshima University Graduate School of Biomedical Science, Hiroshima, Japan. tfuji@hiroshima-u.ac.jp
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+ Y/ z7 z3 {& K0 V' yAbstract! Y: t- r: |5 v& n
BACKGROUND AND OBJECTIVE: Molecular gene markers, which can distinguish human bone marrow mesenchymal stem cells from human fibroblasts, have recently been reported. Messenger RNA levels of tissue factor pathway inhibitor-2, major histocompatibility complex-DR-alpha, major histocompatibility complex-DR-beta, and neuroserpin are higher in human bone marrow mesenchymal stem cells than in human fibroblasts. However, human bone marrow mesenchymal stem cells express less apolipoprotein D mRNA than human fibroblasts. Periodontal ligament cells are a heterogeneous cell population including fibroblasts, mesenchymal stem cells, and progenitor cells of osteoblasts or cementoblasts. The use of molecular markers that distinguish human bone marrow mesenchymal stem cells from human fibroblasts may provide insight into the characteristics of human periodontal ligament cells. In this study, we compared the molecular markers of human periodontal ligament cells with those of human bone marrow mesenchymal stem cells and human gingival fibroblasts. MATERIAL AND METHODS: The mRNA expression of the molecular gene markers was analyzed using real-time polymerase chain reaction. Statistical differences were determined with the two-sided Mann-Whitney U-test. RESULTS: Messenger RNA levels of major histocompatibility complex-DR-alpha and major histocompatibility complex-DR-beta were lower and higher, respectively, in human periodontal ligament cells than in human bone marrow mesenchymal stem cells or human gingival fibroblasts. Human periodontal ligament cells showed the lowest apolipoprotein D mRNA levels among the three types of cells. CONCLUSION: Human periodontal ligament cells may be distinguished from human bone marrow mesenchymal stem cells and human gingival fibroblasts by the genes for apolipoprotein D, major histocompatibility complex-DR-alpha, and major histocompatibility complex-DR-beta.
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, m8 V$ \  [% e3 A1 [8 MPMID: 17451549 [PubMed - indexed for MEDLINE]9 a. W6 V% F; X2 M
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Publication Types, MeSH Terms, SubstancesPublication Types:
7 N8 O0 d- \% K, ~* LComparative Study
, Y( l; m7 i( g5 ^. JMeSH Terms:
# k- C5 o- h2 N8 K) i) K+ YApolipoproteins D/genetics
: p5 I0 c6 j2 F0 H/ t* @" YBone Marrow Cells/cytology
  Z( m' G8 W+ @6 TFibroblasts/cytology*
/ q: t& X9 s8 i& n& LGenetic Markers/genetics
! p' O/ @4 b2 MGingiva/cytology*
1 n. k! N9 T- q* g. w& d8 i3 wHumans
  S9 Y$ W6 x% o( d  e3 OMajor Histocompatibility Complex/genetics) F. E" q( f2 P5 M0 U
Mesenchymal Stem Cells/cytology*& a3 O% H8 v8 t2 N" D- L' p8 Q% D3 s. J
Periodontal Ligament/cytology*: d& {- v) n5 W
RNA, Messenger/analysis# \0 a0 F. Q* I5 n! k* Y1 J* F9 A
Statistics, Nonparametric- j( i6 t4 t8 Z: v, Y( h
Substances:' C9 c7 A/ _3 T: ]2 `
Apolipoproteins D( s* X  m: Q4 ^/ S
Genetic Markers
- M( @2 @& F: K# B6 x$ E) }RNA, Messenger, v2 H! r* i" ?1 F- h; E3 [# G) {9 Z: h
LinkOut - more resourcesFull Text Sources:" V2 f+ G3 _" c- ]. m1 y6 f
Blackwell Publishing
% T+ n) n9 [# L+ X! D" nEBSCO+ U4 u/ V" m  V; O% a
Ingenta plc
: d: b/ \; A; H8 Y6 U& yOhioLINK Electronic Journal Center0 a- h4 w: I5 n/ R" t+ p- {
Ovid Technologies, Inc.
6 k; v: F+ Z" {4 U5 xSwets Information Services# y/ C+ C; t* E5 F; g' a
Medical:
+ c" }2 Q8 d6 k6 CGenetics Home Reference - SERPINI1 Gene - Genetics Home Reference
$ ~1 V4 U, F* Q  A" v6 L& S2.Biochem Biophys Res Commun. 2005 Jun 24;332(1):297-303.' m7 D+ P! D2 R( e0 o! l# I
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Molecular markers distinguish bone marrow mesenchymal stem cells from fibroblasts.
! _  i! ~) f% B5 X3 y) ]' t; Z: K2 O) A$ JIshii M, Koike C, Igarashi A, Yamanaka K, Pan H, Higashi Y, Kawaguchi H, Sugiyama M, Kamata N, Iwata T, Matsubara T, Nakamura K, Kurihara H, Tsuji K, Kato Y.
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Department of Dental and Medical Biochemistry, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.* f/ x2 _; W" x/ ~# [! Y( p1 V4 K
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Abstract
  J4 ^1 I1 x$ g/ QTo characterize mesenchymal stem cells (MSC), we compared gene expression profiles in human bone marrow MSC (11 lines) and human fibroblasts (4 lines) by RT-PCR and real time PCR. Messenger RNA levels of MHC-DR-alpha, MHC-DR-beta, MHC-DR-associated protein CD74, tissue factor pathway inhibitor-2, and neuroserpin were much higher in MSC than in fibroblasts, even in the presence of large interindividual variations. Those of adrenomedullin, apolipoprotein D, C-type lectin superfamily member-2, collagen type XV alpha1, CUG triplet repeat RNA-binding protein, matrix metalloproteinase-1, protein tyrosine kinase-7, and Sam68-like phosphotyrosine protein/T-STAR were lower in MSC than in fibroblasts. FACS analysis showed that cell surface expression of MHC-DR was also higher in MSC than in fibroblasts. MHC-DR expression decreased after osteogenic differentiation, whereas the expression of adrenomedullin-a potent stimulator of osteoblast activity-along with collagen XV alpha1 and apolipoprotein D increased after osteogenic differentiation. The marker genes identified in this study should be useful for characterization of MSC both in basic and clinical studies.+ m* W" g1 g* ]' _4 W6 a

, ~" H: W  a7 K" `+ F3.Stem Cells Dev. 2010 Jun 7. [Epub ahead of print]
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& M" v5 R! ~) a0 K/ Y$ Q% F9 \Markers distinguishing mesenchymal stem cells from fibroblasts are downregulated with passaging.1 `. x6 t. j* W  `2 {/ u5 W
Halfon S, Abramov N, Grinblat B, Ginis IO.
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) I; a+ N# L+ }7 v( e3 d8 B9 ^5 mTeva Pharmaceutical Industries, Cell Therapy Lab, Rehovot, Israel; as5454@gmail.com.
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5 Q5 r2 D  Z, R  e/ P, E" UAbstract
" t# T2 L6 Y( q) t  rExpansion of plastic-adherent bone marrow-derived MSC results in gradual loss of osteogenic potential after passage 5-6. One explanation is contamination of MSC cultures with mature cells including fibroblasts. Identification and elimination of fibroblasts from MSC cultures could improve MSC yield and differentiation potential and also prevent tumor formation after MSC transplantation. However, no specific markers currently exist that can reliably discriminate MSC from fibroblasts. Flow cytometry analysis demonstrated that markers currently used to define MSC, such as CD105, CD166, CD90, CD44, CD29, CD73 and CD9 are also expressed on human skin or lung fibroblasts. However the level of expression of CD166 was significantly higher and that of CD9 was significantly lower in MSC than in fibroblasts. CD146 was expressed only in MSC. Using small focused microarrays new markers differentially expressed in MSC and fibroblasts were identified. Real Time PCR confirmed that expression of CD106, ITGA11, and IGF-2 in MSC was at least 10 fold higher than in fibroblasts while expression of MMP1 and MMP3 was almost 100 fold lower. Flow cytometry and immunostaining demonstrated that CD106 protein expression on cell surface could be upregulated in MSC but not in fibroblasts by the treatment with TNF-alpha. Comparison of surface expression of commonly used and newly identified MSC markers in MSC cultures of passage 2 and of passage 6 demonstrated that CD106 (with and without TNF-alpha treatment), ITGA11 and CD146 were downregulated in MSC of passage 6 and CD9 was upregulated, while all other markers did not change. Newly identified markers that have robust differences of expression in MSC and fibroblasts on gene and protein level could be used for quality control of MSC cultures after expansion, cryopreservation, gene transfection and other manipulations.

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