[EFFECT OF FETAL BOVINE SERUM ON OSTEOGENIC GROWTH PEPTIDE PROMOTING BONE MARROW MESENCHYMAL STEM CELLS PROLIFERATION AND DIFFERENTIATION].

OBJECTIVE

To explore the effect of fetal bovine serum (FBS) of different concentrations in the culture medium on osteogenic growth peptide (OGP) promoting bone marrow mesenchymal stem cells (BMSCs) proliferation and differentiation.

METHODS

BMSCs were separated from limb bones of 8 Sprague Dawley rats (5 weeks old) and purified by adherence method, and BMSCs at passage 3 were divided into 4 groups according to OGP concentration: OGP 1 x 10(-10) mol/L group, OGP 1 x 10(-9) mol/L group, OGP 1 x 10(-8) mol/L group, and control group without OGP; and 0, 2%, 5%, 8%, and 10% FBS concentration gradient was used in each group. The cell proliferation rate was detected by MTT method at 1, 3, 5, 7, 9, and 12 days after culture, and the activity of intracellular alkaline phosphatase (ALP) was determined by the method of p-nitrophenyl phosphate disodium at 9 days after culture.

RESULTS

BMSCs showed adherent growth, rapid proliferation, long fiber vortex, and typical morphology. MTT analysis showed that cells could not sustain proliferation when FBS concentration was less than 5% in each group; when FBS concentration was above 8%, cells proliferated continually. Proliferation promoting effect of OGP 1 x 10(-8) mol/L and 1 x 10(-9) mol/L groups was significantly higher than that of the control group in all serum concentrations (P < 0.05); when FBS concentration was lower than 10%, the proliferation promoting effect of OGP 1 x 10(-8) mol/L group was significantly higher than that of the other 2 OGP groups (P < 0.05), but when FBS concentration was 10%, OGP 1 x 10(-8) mol/L group had no advantage of promoting proliferation. ALP test results showed that as the FBS concentration increased, ALP activity of all groups also significantly increased (P < 0.05). Under the condition of 5% FBS and 8% FBS, the ALP activity of each OGP group was significantly greater than that of the control group, and it was the highest in OGP 1 x 10(-8) mol/L group (P < 0.05). Under the condition of 10% FBS, the ALP activity of each OGP group was still greater than that of the control group (P < 0.05), but no significant difference was found between the OGP 1 x 10(-8) mol/L group and OGP 1 x 10(-9) mol/L group (P > 0.05).

CONCLUSION

The concentration of 8% FBS is the best concentration of serum for OGP promoting the proliferation and differentiation of BMSCs, and the most suitable concentration of promoting the proliferation and differentiation of BMSCs is OGP 1 x 10(-8) mol/L.