Tonsil-derived mesenchymal stromal cells: evaluation of biologic, immunologic and genetic factors for successful banking.
OBJECTIVE
Although mesenchymal stromal cells (MSC) from human palatine tonsils (tonsillar MSC, T-MSC) have been isolated, whether T-MSC isolated from multiple donors are feasible for cell banking has not been studied.


METHODS
T-MSC before and after a standard protocol of cryopreservation and thawing were assessed regarding several basic characteristics, including colony-forming unit-fibroblast features, MSC-specific surface antigen profiles, and inhibition of alloreactive T-cell proliferation. In vitro mesodermal differentiation potentials to adipocytes, osteocytes and chondrocytes were detected by staining with either cell-specific dyes or antibody after incubation with each appropriate differentiation medium. Expression of mesoderm-specific genes was also quantified by real-time polymerase chain reaction (PCR) assay. Expression profiles of endoderm-specific genes were identified by reverse transcription PCR assay. The feasibility of T-MSC in future engraftment was tested by short tandem repeat (STR) analysis using genomic DNA isolated randomly from three independent subjects.


RESULTS
Both fresh and cryopreserved-thawed T-MSC showed a similar high proliferation capacity and expressed primitive cell-surface markers. Hematopoietic cell markers, HLA-DR, co-stimulatory molecules and follicular dendritic cell markers were not detected. In addition to mesodermal differentiation, fresh and cryopreserved-thawed cells also underwent endodermal differentiation, as evidenced by the expression of endoderm-specific genes including forkhead box A2 (FoxA2), SIX homeobox 1 (Six1) and chemokine (C-C motif) ligand 21 (CCL21). Both cells significantly decreased phorbol 12- myristate 13-acetate (PMA)-induced T-cell proliferation. T-MSC from three independent donors formed chimerism in STR analysis.


CONCLUSIONS
Our results demonstrate for the first time that T-MSC are a potentially good source for MSC banking.
Department of Pediatrics, School of Medicine, Ewha Womans University, Seoul, Korea.
Kyung-Ha
KH
Ryu
Kyung-Ah
KA
Cho
Hae Sang
HS
Park
Ji-Yon
JY
Kim
So-Youn
SY
Woo
Inho
I
Jo
Yoon Hee
YH
Choi
Young Mi
YM
Park
Sung-Chul
SC
Jung
Sung Min
SM
Chung
Byung-Ok
BO
Choi
Han Su
HS
Kim
D000954:Antigens, Surface / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D046528:Chimerism; D003114:Colony-Forming Units Assay; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism; D014066:Palatine Tonsil / Q000166:cytology*; D010641:Phenotype; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D014015:Tissue Banks; D014019:Tissue Donors
D000954:Antigens, Surface