Derivation and Characterization of hMSCs from hESCs

hMSCs were differentiated from hESCs based on a published protocol^{31 (link)}. Briefly, embryoid bodies were left to differentiate in αMEM (Invitrogen) medium supplemented with 10% FBS (AusGeneX), 10 ng/ml bFGF (JPC), 5 ng/ml TGFβ (HumanZyme) and 1% penicillin/streptomycin (Gibco) until fibroblast-like cells appeared. The hMSCs were purified with different antibodies corresponding to hMSC-specific markers (CD73, CD90, and CD105) by FACS. Antibodies used for hMSC characterization were as follows: anti-CD105-APC (17-1057-42) antibody was purchased from eBioscience; anti-CD90-FITC (555595), anti-CD73-PE (550257), anti-CD34-PE (555822), anti-CD43-APC (580198), and anti-CD45-FITC (555482) antibodies were purchased from BD Biosciences. Anti-IgG-FITC (555748), anti-IgG-PE (555749), and anti-IgG-APC (555751) antibodies from BD Biosciences were used as isotype controls. The functionality of hMSC was further verified by differentiation towards cartilage, bone, and adipocytes^{31 (link)}. The tri-lineage differentiation abilities of hMSC lines were evaluated by histochemical staining with von Kossa (osteogenesis), Alcian blue (chondrogenesis), and Oil red O (adiopogenesis) Kit (IHC World), respectively.

Free full text: Click here

Pan H., Guan D., Liu X., Li J., Wang L., Wu J., Zhou J., Zhang W., Ren R., Zhang W., Li Y., Yang J., Hao Y., Yuan T., Yuan G., Wang H., Ju Z., Mao Z., Li J., Qu J., Tang F, & Liu G.H. (2016). SIRT6 safeguards human mesenchymal stem cells from oxidative stress by coactivating NRF2. Cell Research, 26(2), 190-205.

Publication 2016

αMEM penicillin/streptomycin Alcian blue Oil red O (adiopogenesis) Kit

Alcian blue Anti igg Antibodies Antibody Bone Cartilage Cd43 Cd73 Cd90 Chondrogenesis Embryoid bodies Fibroblast cells Fitc Hescs Isotype Oil red o Osteogenesis Penicillin Streptomycin Tgfβ

Corresponding Organization : Ministry of Education of the People's Republic of China

Other organizations : Institute of Biophysics, Chinese Academy of Sciences, Peking University, Salk Institute for Biological Studies, University of Iowa, Foshan University, Hangzhou Normal University, Tongji University, Beijing Hospital, Institute of Zoology

Top 5 similar protocols

Protocol cited in 10 other protocols

Variable analysis

independent variables

HESC differentiation protocol
Differentiation factors (10% FBS, 10 ng/ml bFGF, 5 ng/ml TGFβ)

dependent variables

Appearance of fibroblast-like cells
Expression of hMSC-specific markers (CD73, CD90, CD105)
Tri-lineage differentiation ability (cartilage, bone, adipocytes)

control variables

Cell culture medium (αMEM)
Penicillin/streptomycin (1%)
Isotype control antibodies (anti-IgG-FITC, anti-IgG-PE, anti-IgG-APC)

controls

Positive control: Tri-lineage differentiation ability of hMSCs
Negative control: Isotype control antibodies for hMSC characterization

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!