Identification of Bone Marrow Stromal Cells and
Expression of Tyrosine Hydroxylase Gene in Them
LU
Ling-Ling, SU Yu-Jin, ZHAO Chun-Li, ZOU Xi-Feng, HOU Shao-Ping, XU Qun-Yuan, YANG Hui*
( Beijing
Institute for Neuroscience, Capital University of Medical Sciences, Beijing
100054, China )
Abstract The
study is to establish the method of isolation and identification of bone marrow
stromal cells and to investigate the ability of bone marrow stromal cells to
accept and express TH gene. Cells were isolated by a density gradient
(lymphocytes separation) and identified by BrdU labeling and
fluorescence-activated cell sorting (FACS) technology using CD11b, CD45 and
CD90 antibodies. TH and lacZ gene were transfected to rBMSCs with an
adeno-associated virus vector. The results showed that most tightly adherent
cells in the primary culture were fibroblast-like and formed foci of two to
four cells. The cells in the foci remained dormant for 2 to 4 days and then
began to multiply rapidly. After several passages, the adherent cells became
more uniformly spindle-shaped in appearance. BrdU, indicating that BMSCs
replicate actively, labeled about 74.9% of cultured cells. Data from FACS
showed that about 75% of isolated cells were CD90+/CD45–/CD11b–, which is
the marker of bone marrow stromal cells. The efficiency of TH gene transfection
was about 75%. BMSCs could readily be genetically engineered and could be
useful delivery targets of gene therapy for Parkinson’s disease.
Key words bone
marrow stromal cells; Parkinson’s disease; adeno-associated virus vector; TH
gene
*Correspondence: Tel, 86-10-63051480; Fax, 86-10-63056992; e-mail,
[email protected]