Basic biological characteristics of adipose mesenchymal stem cells and the characteristics in differentiation into osteoblast

Xiaodong JU; Siquan LOU; Hua TIAN; Weiguo WANG; Yanqing LIU

ABSTRACT

BACKGROUND:

Bone marrow mesenchymal stem cells(MSCs) have been investigated initially. Because both fat tissue and bone marrow are tissues originated from mesoderm, so whether MSCs could be obtained from fat tissue as well, which also have multi-lineage differentiation potential?

OBJECTIVE:

To investigate the basic biological characteristics of adipose MSC and its differentiation into osteoblast under given culture condition for the exploration of its feasibility as seed cell in bone tissue engineering.

DESIGN:

A randomized and controlled study based on cells.

SETTING:

Department of Orthopaedics of an Affiliated Hospital of a university.MATERIALS The study was conducted in the Department of Orthopaedics of the Third Hospital of Peking University. Adipose MSC extracted from the fat tissue of Lewis rats was used as subject.

METHODS:

Adipose MSCs were obtained from inguinal fat pads of Lewis rat after digestion, which were induced into adipocytes and osteoblasts with adipose and osteogenesis induced culture mediums. The differentiations were examined with cytochemical staining, immuncytochemical staining and Western blotting.MAIN OUTCOME

MEASURES:

Morphological and biological characteristic of adipose MSCs, and the specific mark of osteoblast after induction.

RESULTS:

Adipose MSCs were obtained from rat adipose tissue culture,which appeared fibroblast-likely in the culture in vitro, and could stably proliferate and passage in vitro. Primary adipose MSCs could differentiate into adipocytes spontaneously, and passaged cells could form fat drop under the reaction of insulin and dexamethasone, and then differentiate towards adipocytes after the expression of peroxidase proliferation activated receptor ?(PPAR-?) enhanced. There was significant difference between induction group and control group in alkaline phosphatase(ALP) activity detection under the induction of dexamethasone, ascorbic acid, and ?-sodium glycerophosphate(P ＜ 0.01) . Calcium node appeared in yon Kossa staining. Result of osteopontin (OPN) immunocytochemcial staining was positive,and OPN expression was detected by Western blotting after induction.

CONCLUSION:

MSCs with multi-lineage potential can be obtained from rat adipose tissue, and differentiate into adipocytes and osteoblasts after inductions. Therefore, adipose MSCs can possibly be served as one of optimal seed cells for bone tissue engineering.