Abstract
Background: Expanding cells ex vivo is an
important step in tissue engineering and the culture
medium is usually supplemented with bovine
serum. When a patient receives bone marrow
stromal cells (BMSCs) grown in a medium containing
bovine serum, infection of bovine diseases
and/or the patient’s unfavorable immune reaction to
bovine proteins are of concern. To overcome
these problems, we examined whether a patient’s
autologous serum could support the growth and
differentiation of his/her BMSCs.
Methods: Bone marrow was collected from the
iliac by aspiration and cultured in a medium supplemented
with 10% autologous serum or 10%
fetal bovine serum (FBS). Number and area of the
colonies of fibroblast-like cells (colony-forming
unit fibroblast, CFU-f) were measured 8 days after
the cell inoculation (day 8). Number and area of the
alkaline phosphatase (ALP) positive colonies
were measured on day 10. On day 40, the cultures
were stained with alizarin red S. RNA was prepared
from the culture on day 20, and the mRNA expression
of osteoblastic genes was examined by
reverse transcription polymerase chain reaction
(RT-PCR) analysis.
Results: BMSCs, which were cultured in the
medium supplemented with autologous serum,
produced CFU-f, ALP-positive area and mineralized
nodules, which is comparable to the BMSCs in the
culture supplemented with FBS. The mRNA
expressions of osteopontin, parathyroid hormone
receptor, osteocalcin, and bone sialoprotein were
detected in the culture supplemented with autologous
serum.
Conclusions: A patient’s autologous serum
could expand BMSCs without losing their potentiality
for osteoblastic differentiation. Patients'
autologous serum could be efficient to expand
BMSCs and to be utilized safely for bone regeneration
therapy.
