Bone marrow mesenchymal stem cells cultured by umbilical cord blood serum versus adult autoserum in vitro

Wenyong KUANG; Xinfu ZHOU; Ruijuan LI; Mincui ZHENG

ABSTRACT

BACKGROUND:

Studies concerning bone marrow mesenchymal stem cells (BMSCs) cultured by umbilical cord serum in vitro were hot topic to avoid the heterogenous serum rejection during BMSC culture and improve culture efficiency of BMSCs.

OBJECTIVE:

To compare biological feature of BMSCs by the umbilical cord serum and adult autoserum in vitro.DESIGN,TIME AND

SETTING:

The opening experiment was performed at the Laboratory of Cell Biology,Xiangya Second Hospital,Central South University,China from October 2006 to June 2008.MATERIALSSixteen bone marrow samples were provided by Xiangya Second Hospital,Central South University and Xiangtan Central Hospital of Hunan Province.

METHODS:

BMSCs were obtained from 16 fresh adult bone marrow by gradient centrifugation with Ficoll Paque,cultured with DMEM/F12 with 10% umbilical cord blood serum and 10% adult autoserum.The fourth passage was used in this study.The surface antigens of BMSCs were detected by flow cytometry.BMSCs could differentiate into ostenblasts and adipocytes under culture in conditioned medium for osteogeneais and adipogenesis and the differentiated BMSCs were identified by morphological observation,immunophenotype and immunochemical staining.MAIN OUTCOME

MEASURES:

The following parameters were measuredmorphological observation;cell cycle and cell count;identification of surface antigen;observation of osteoblasts and adipocytea induced from BMSCs by immunochemical staining.

RESULTS:

The quantity and velocity of BMSCs in umbilical curd blood serum was better than those in adult autoserum (P ＜ 0.05)Only few cells were proliferating in both medium,BMSCs in S phase in umbilical cord blood serum was more than those in adult autoserum (P ＜ 0.05).The positive rate of CD29,CD73 and CD105 on BMSCs in umbilical cord serum (above 90%) was higher than those in adult autoserum (P ＜ 0.05),and the positive rates of CD31,CD34 and CD45 were lower than 2%,and the positive rate of CD31 was lower than those in adult autoserum (P ＜ 0.05).The positive rate of BMSCs differentiated into osteoblasts and adipocytes in umbilical cord blood serum was also higher than these in adult autoserum (P ＜ 0.05).

CONCLUSION:

Purity and differentiated potency of BMSCs in umbilical cord blood serum are better than those in the adult autoserum in vitro.The umbilical cord serum is more adapt to clinical application.