[Isolation and identification of chronic myelogenous leukemia bone marrow mesenchymal stem cells and their functional characteristics].
OBJECTIVE
To isolate and culture bone marrow mesenchymal stem cells (MSCs) from chronic myelogenous leukemia (CML) patients and examine their functional characteristics.


METHODS
Bone marrow was extracted from the anterior superior iliac spines of 21 patients with CML. MSCs were isolated and cultured. Single colony derived MSCs were harvested by limiting dilution. The cell cycle and immunophenotype of the expanded clonal MSCs were detected by fluorescence-activated cell sorter (FACS). Different agents were used to induce the MSCs to differentiate into osteocyte, adipocyte and neural cells. Von Kossa staining, oil-red staining, and Western blotting was used to examine the ability of differentiation. PCR was used to detect the expression of BCR/ABL gene. The ultrastructure of the CML derived MSCs was observed with electron microscopy. Sixteen BALB/c nude mice were randomly divided into 2 equal groups to be inoculated with HL60 cancer cells and CML derived MSCs to observe the tumorigenicity. MSCs were cultured in soft agar for 2 weeks to observe the clone growth.


RESULTS
Fibroblast-like, positive in CD29, CD44, and CD105, and negative in CD116, CD34, CD48, and HLA-DR, the CML derived MSCs could differentiate into osteocyte, adipocyte and neural cells. CML derived MSCs showed normal karyotype and ultrastructure, they did not express BCR/ABL gene. After 2 weeks' culture no clone was formed from the MSCs. Four weeks after tumors were shown in 6 of the 8 mice inoculated with HL60 cells, and no tumor was seen in the mice inoculated with MSCs.


CONCLUSIONS
Able to differentiate into different types of cell and without tumorigenicity, MSCs from the bone marrow of CML patients have the potentiality in clinical application.
Zhao
Zhi-gang
ZG
Department of Hematology, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, China.
Tang
Xiao-qiong
XQ
Li
Jing
J
Shi
Ming-xia
MX
Zou
Ping
P
D000818:Animals; D001854:Bone Marrow Cells / Q000473:pathology* / Q000502:physiology; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000473:pathology* / Q000601:surgery; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000473:pathology* / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D011897:Random Allocation