The mesenchymal stem cell antigen MSCA-1 is identical to tissue non-specific alkaline phosphatase.
We have recently identified 2 distinct CD271(bright)MSCA-1(dim)CD56(+) and CD271(bright)MSCA-1(bright)CD56(-) MSC subsets in primary femur-derived bone marrow (BM), which differ in their expression pattern and morphology as well as in their clonogenic and differentiation capacity. Here, we show that MSCA-1 is identical to tissue non-specific alkaline phosphatase (TNAP), an ectoenzyme known to be expressed at high levels in liver, bone, and kidney as well as in embryonic stem (ES) cells. SDS-PAGE of WERI-RB-1 cell lysate and supernatant from phosphatidylinositol-specific phospholipase C (PI-PLC)-treated WERI-RB-1 cells resulted in the appearance of a prominent 68-kDa band. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDITOF MS) sequence analysis revealed TNAP-specific peptides. Screening of the MSCA-1-specific antibody W8B2 on HEK-293 cells transfected with the full-length coding sequence of TNAP showed specific reactivity with transfected but not with parent cell line. In addition, TNAP-specific mRNA expression was selectively detected in the transfectant line. In agreement with these findings, enzymatic activity of TNAP was exclusively detected in sorted MSCA-1(+) BM cells but not in the MSCA-1(-) negative fraction. Surface marker analysis revealed coexpression of the embryonic marker SSEA-3 but not SSEA-4, TRA-1-60, and TRA-1-81. In endometrium, TNAP is expressed at intermediate levels on CD146(+) cells and at high levels in the luminal space of glandular epithelia. Our results demonstrate that TNAP is a selective marker for the prospective isolation of BM-derived MSC and MSC-like cells in endometrium.
Sobiesiak
Malgorzata
M
Department of Internal Medicine II, Division of Hematology, Immunology, Oncology and Rheumatology, University Clinic of Tübingen, Tübingen, Germany.
Sivasubramaniyan
Kavitha
K
Hermann
Clemens
C
Tan
Charmaine
C
Orgel
Melanie
M
Treml
Sabrina
S
Cerabona
Flavianna
F
de Zwart
Peter
P
Ochs
Uwe
U
Müller
Claudia A
CA
Gargett
Caroline E
CE
Kalbacher
Hubert
H
Bühring
Hans-Jörg
HJ
D000328:Adult; D000469:Alkaline Phosphatase; D000954:Antigens, Surface / Q000032:analysis / Q000302:isolation & purification; D015415:Biomarkers; D001854:Bone Marrow Cells / Q000166:cytology; D002352:Carrier Proteins / Q000032:analysis / Q000302:isolation & purification; D004717:Endometrium / Q000166:cytology; D005260:Female; D006801:Humans; D008875:Middle Aged; D014018:Tissue Distribution
D000954:Antigens, Surface; D015415:Biomarkers; D002352:Carrier Proteins; C540400:W8B2 protein, human; C548267:ALPL protein, human; D000469:Alkaline Phosphatase