1 骨髓间充质干细胞的发现和来源
骨髓组织中有多种细胞成分，除基质细胞等已经分化的细胞外，还含有两类多潜能干细胞：造血干细胞和间充质干细胞。1987 年Friedenstein 等发现在塑料培养皿中培养的贴壁的骨髓单个细胞在一定条件下可分化为多种类型的细胞，而且经过20－30个培养周期仍能保持其多向分化潜能。由于骨髓中的这种多能细胞能够分化为多种中胚层来源的间质细胞, 故称之为间充质干细胞(Mesenchymal stem cells，MSCs),或间质祖细胞(MPCs)，是成人多能干细胞的一类。早期分离培养时，发现其形状呈成纤维细胞样而称其为成纤维细胞集落形成单位(Colony-forming unit-fibroblast，CFU-F)，或骨髓基质成纤维细胞(Marrow stromal fibroblast，MSF)。Friedenstein AJ , Chailakhyan RK, Gerasimov UV. Bone marrow o steogenic stem cells: in vit ro cult ivat ion and t ransp lantat ion in diffusion chambers. Cell T issue Kinet, 1987, 20 (3) : 263-267]
2 鉴于其强大的增殖能力及多向分化潜能，可在体外长期培养和遗传背景较稳定，而且用自体干细胞诱导构建的组织不涉及伦理问题，也不存在MHC限制，所以骨髓间充质干细胞日益受到重视。但是与造血干细胞等其他细胞相比，骨髓中MSCs的数量非常少，约占整个骨髓有核细胞的十万分之一，并随年龄的增加，细胞数量逐渐减少。因此，如何简便有效地从骨髓中获取高纯度的MSCs显得尤为重要，寻找高度特异性的MSCs的表面抗原也就成为MSCs研究中的一项重要任务和目标。
不仅如此，一种同样来源于骨髓、贴壁生长、被认为更原始（可以分化为MSCs）也具有更强增殖能力的干细胞也被鉴定，它就是多能成体祖细胞（multipotent adult progenitor cell (MAPC) or mesodermal progenitor cell(MPC)）[Reyes, M.,
C.M. (2001) Purification and in vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98, 2615-2625]，因能和MSCs一起被纯化而统称BM stromal stem cell。
3 利用流式细胞仪的研究显示,MSCs属混杂细胞群，其表面抗原也具有非专一性, 它表达了间质细胞、内皮细胞和表皮细胞的表面标志。主要包括: ①黏附分子, 如CD166、CD54、CD102、CD44、CD106 等。②生长因子和细胞因子受体, 如白介素21 受体( IL-1R )、IL-3R、IL-4R、IL-6R、IL-7R、C干扰素受体( IFN-CR)、肿瘤坏死因子(TNF) -A等。③整合素家族成员, 包括CD
但到目前为止, 对于MSC 的表面标志尚不完全确定，有些标志尚存在种属特异性，现将普遍认同的不同种属的MSCs的表面抗原情况综述如下：
表一：MSCs的表面抗原
|
HUMAN |
RAT |
MOUSE |
REFERENCE |
|
SH2 +and SH3+ |
|
|
Caplan AI. Mesenchymal stem cells. J.Orthop.Res. 1991;9:641-650. |
|
CD105+ |
|
|
Barry FP, et al. The monoclonal antibody SH-2, raised against human mesenchymal stem cells, recognizes an epitope on endoglin (CD105). Biochem.Biophys.Res.Commun. 1999;265:134-139. |
|
CD73+ |
|
|
Barry F, Boynton R, Murphy M, Zaia J. The SH-3 and SH-4 Antibodies Recognize Distinct Epitopes on CD73 from Human Mesenchymal Stem Cells. Biochemical and Biophysical Research Communications 2001;289:519-524. |
|
SH2+, SH3+, CD29+, CD44+, CD71+, CD90+,CD106+, CD 脂多糖受体CD14-、CD34- CD45-。 |
|
|
Dean RJ. Moseley AB. Mesenchymal stem cells: biology and potential clinical uses. Experim Hematol, 2000,28: 875-884 |
|
CD105+, CD73 +,CD90+; CD45-, CD34-, CD14- or CD11b-, CD or CD19- and HLA class II-. |
|
|
M Dominici1,et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy.2006,8( 4):315-317 |
|
CD29+ and CD44+, but CD34_ and CD45_ |
|
|
Hou M, et al.Transplantation of mesenchymal stem cells from human bone marrow improves damaged heart function in rats. Int J Cardiol. 2007 ,115(2):220-8.. |
|
the neural ganglioside GD2+ |
|
|
Human Bone Marrow Mesenchymal Stromal Cells Express the Neural Ganglioside GD2: A Novel Surface Marker for the Identification of MSCs. Blood,2006 |
|
STRO-1+ |
|
|
Encina, N.R., et al . Immunomagnetic isolation of osteoprogenitors from human bone marrow stroma. Lab. Invest. 1999, 79, 449-457. |
|
NGFR+ (nerve growth factor receptor ) |
|
|
N. Querici, et al . Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies, Exp. Hematol. 30 (2002)783–791 |
|
CD11b- (C57Bl/6 mice) |
|
|
Philippe Tropel, et al. Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow. Experimental Cell Research 295 (2004) 395– 406 |
|
CD105+ |
|
|
Barry FP, Boynton RE, Haynesworth S, Murphy JM, Zaia J. The monoclonal antibody SH-2, raised against human mesenchymal stem cells, recognizes an epitope on endoglin (CD105). Biochem.Biophys.Res.Commun. 1999; 265: 134-139. |
|
CD73+ |
|
|
Barry F, Boynton R, Murphy M, Zaia J. The SH-3 and SH-4 Antibodies Recognize Distinct Epitopes on CD73 from Human Mesenchymal Stem Cells. Biochemical and Biophysical Research Communications 2001;289:519-524. |
|
CD271+ （But it is lost during culture） |
|
|
Jones EA, et al. Isolation and characterization of bone marrow multipotential mesenchymal progenitor cells. Arthritis Rheum. 2002;46:3349-3360. |
|
D7Fib+ （It is also expressed on skin fibroblasts.） |
|
|
Jones EA, et al. Enumeration and phenotypic characterization of synovial fluid multipotential mesenchymal progenitor cells in inflammatory and degenerative arthritis. Arthritis Rheum. 2004;50:817-827. |
|
SH2 (scr同源区2) +, SH3+ CD105+ CD73+ |
|
|
Dominici M, Le BK, Mueller I et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315-317. |
|
SH-2+ , B-10+,CD14-,CD45-(MSCs); CD14-,CD45- ( marrow-derived stromal cells, MDSCs) |
|
|
Majumdar MK,et al. Phenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells. J Cell Physiol ,1998,176:57-60 |
|
SH2+, SH3+, CD29+,CD44+, CD71+, CD90+, CD106+, CD lipopolysaccharide receptor-,CD14-, CD34-, and the leukocyte common antigenCD45- |
|
|
Pittenger MF. Multilineage potential of adult human mesenchymal stem cells.science,1999;284:143-146 |
|
CD1α-,CD14-, CD31-, CD34-,CD56- 、CD45-,上皮特异性肌动蛋白( ESA)- |
|
|
Conget PA ,Minguell JJ . Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol ,1999 ,181 (1) :67-73. |
|
CD29+, CD44+; CD34- CD45- |
|
|
Mai Hou, et al. Transplantation of mesenchymal stem cells from human bone marrow improves damaged heart function in rats. International Journal of Cardiology, 115 (2007) 220-228 |
|
I-HLA+, HLA-DR+, CD44+ |
|
|
An Van Damme, et al. Bone Marrow Stromal Cells as Targets for Gene Therapy. Current Gene Therapy, 2002, 2, 195-209 |
|
CD45-，CD146+，CD117+，Stro-1+ |
|
|
Shi S, Gronthos S. Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res 2003; 18:696-704. |
|
CD (α1-integrin Subunit ) |
CD (rats strain LEW/CRL ICO) |
CD (Mice,strains BALB/cByJ H-2d,C57BL/6J, FVB/N) |
Gindraux F, et al. Human and rodent bone marrow mesenchymal stem cells that express primitive stem cell markers can be directly enriched by using the CD |
|
|
（different rat）CD90+, CD44+, CD54+, CD 106+, CD45-, CD11b- |
|
Heterogeneity of stromal precursor cells isolated from rat bone marrow. Tsitologiia. 2007;49(1):40-47 |
|
|
CD11b (-), CD45 (-), and CD90 (+). (Sprague-Dawley rats) |
|
Yoshimura H, et al. Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Res. 2007 Mar;327(3):449-62. Epub 2006 Oct 13. |
|
|
Vimention+, fibronectin+ (Sprague-Dawley rats) |
|
Pang-Yen Tseng, et al. Spontaneous differentiation of adult rat marrow stromal cells in a long-term culture. J Vet. Med. Sci. 69(2): 95-102,2007. |
|
|
CD11b (-), CD45 (-), and CD90 (+). (Sprague-Dawley rats) |
|
Hideya Yoshimura. Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Res (2007) 327:449–462 |
|
|
CD44+, CD106+, and CD117+ , CD31- (Sprague-Dawley rats) |
|
Jun Zhang, et al. Engineering of Vascular GraftsWith Genetically Modified Bone Marrow Mesenchymal Stem Cells on Poly (Propylene Carbonate) Graft. Artif Organs, 30(12) :898–905, 2006 |
|
|
CD44+ , CD90+, CD45- (Wistar rats ) |
|
Alexandra Stolzing, et al. Stressed Stem Cells: Temperature Response in Aged Mesenchymal Stem Cells. Stem Cell and Development, 15:478–487 (2006) |
|
|
CD34-, CD45-, CD44+ , and SH3+ (Wistar rats) |
|
Yang JF,et al.Transfection of human VEGF165 gene into bone marrow mesenchymal stem cells in rats. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2006 Jun;31(3):313-8. |
|
|
CD29 + ,CD34 low/- (Rat BMSCs) |
|
XiaoHong Li, et al. Bone marrow mesenchymal stem cells differentiate into functional cardiac phenotypes by cardiac microenvironment. Journal of Molecular and Cellular Cardiology 42 (2007) 295-303 |
|
|
|
（Murine） CD140b (PDGFRβ) |
Yoko Koide, et al. Two Distinct Stem Cell Lineages in Murine Bone Marrow .Stem Cells, 2007;25:1213–1221 |
表二：multipotent adult progenitor cells ( MAPCs) 的表面抗原
|
HUMAN |
RAT |
MOUSE |
REFERENCE |
|
I and II MHC-, CD34-, CD44-, CD45-, c-Kit-, Flk-1low,Sca-1 low and Thy-1 low CD13high, SSEA-I high |
CD45-TER119- |