Protocols for cloning human bone marrow-derived hepatic stem cells in vitro.
- Author:
Jie YUAN
1
;
Cai-xian LIAO
;
An-cheng QIN
;
Xin-xin LIAO
;
Yong-ping HUANG
;
Zu-yuan GONG
;
Hui LIAO
Author Information
- Publication Type:Journal Article
- MeSH: Bone Marrow Cells; cytology; Cell Culture Techniques; Clone Cells; Hepatocyte Growth Factor; pharmacology; Hepatocytes; cytology; physiology; Humans; Liver; cytology; Proto-Oncogene Proteins c-kit; metabolism; Stem Cells; cytology; Thrombopoietin; pharmacology
- From: Journal of Southern Medical University 2010;30(2):318-320
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore practical protocols for cloning bone marrow-derived hepatic stem cells in vitro.
METHODSThe cell fraction rich in CD117(+) cells and CD184(+) cells was separated from fresh bone marrow by density gradient centrifugation and cultured for 0, 7 and 14 days in high-glucose DMEM supplemented with or without 10% autologous serum or in serum-free high-glucose DMEM. All the media were supplemented with different concentrations of hepatocyte growth promoting factors (HGPF), thrombopoietin (TPO) and interleukin-3 (IL-3). The quantitative changes of CD117(+) cells and CD184(+) cells were measured by flow cytometry.
RESULTSThe optimal effect for cell cloning was achieved with high-glucose DMEM with 10% autologous serum group supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3. At day 7 of cell culture in this media, the quantity of CD117(+) cells and CD184(+) cells increased by 6.55 and 6.20 folds, and by 11.62 and 20.57 folds at day 14, respectively.
CONCLUSIONIt is practical for cloning bone marrow-derived hepatic stem cells in high-glucose DMEM with 10% autologous serum supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3.
