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

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