Establishment and characterization of a human telomerase catalytic subunit-transduced fetal bone marrow-derived osteoblastic cell line.
The fate of human hematopoietic stem cells (HSCs)/progenitor cells (HPCs) is influenced by bone marrow (BM) stromal cells. To investigate the role of stromal cells in the hematopoietic support, we have transduced human fetal BM stromal cells (FBMSCs) with a human telomerase catalytic subunit (hTERT). One of the resultant cell lines was identified as osteoblasts, because it contained mineral deposits and constitutively expressed osteogenic genes osteocalcin, osteopontin, collagen type I, osteoblast marker alkaline phosphatase, but not marrow stromal cell marker STRO-1 and CD105. The hTERT-transduced fetal BM-derived osteoblastic cells (FBMOB-hTERT) can actively maintain the capacity of self-renewal and multipotency of HSCs/HPCs at least partly through transcriptional up-regulation of hematopoietic growth factors such as stem cell growth factors (SCFs) and Wnt-5A during interaction with HSCs/HPCs. The enhanced transcription of SCFs and Wnt-5A appears to be mediated by CD29 signaling. Moreover, the FBMOB-hTERT cells seem superior to primary FBMSCs in supporting hematopoiesis, because they are more potent than primary FBMSCs in supporting the ex vivo expansion and long-term culture initiating cells activity of HSCs. The FBMOB-hTERT cell line has been maintained in vitro more than 125 population doublings without tumorigenicity. The results indicate that the FBMOB-hTERT is useful for the study of molecular mechanisms by which osteoblasts support hematopoiesis.
Yang
Yu-Xia
YX
State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China.
Miao
Zhen-Chuan
ZC
Zhang
Hao-Jian
HJ
Wang
Yun
Y
Gao
Jian-Xin
JX
Feng
Mei-Fu
MF
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000201:enzymology; D020134:Catalytic Domain; D002460:Cell Line / Q000166:cytology* / Q000201:enzymology; D005333:Fetus / Q000166:cytology; D006410:Hematopoiesis / Q000235:genetics; D016298:Hematopoietic Cell Growth Factors / Q000235:genetics / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000201:enzymology; D006801:Humans; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000201:enzymology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000201:enzymology; D019098:Telomerase / Q000235:genetics*; D014161:Transduction, Genetic; D015854:Up-Regulation; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism; D000071818:Wnt-5a Protein; D019061:src-Family Kinases
D016298:Hematopoietic Cell Growth Factors; D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D019061:src-Family Kinases; C509186:TERT protein, human; D019098:Telomerase