We screened the differentiation-inducing activities of 39 mushroom extracts from Akita prefecture, Japan, on the mouse osteoblastic cell line, MC
3
T
3
-
E
1. Sixteen phosphate buffered saline (PBS), 8 boiled PBS, 14 ethanol and 12 methanol extracts induced alkaline phosphatase (ALP) activities, an indicator of MC
3
T
3
-
E
1 cell differentiation. The enzyme activities were markedly induced by extracts of
Tricholoma auratum, and we isolated the active compound from methanol extracts of this mushroom. Physical data for the isolated active compound were identical to those for (
22E
,24
R)-ergosta-7,
22
-diene-
3β
,
5α
,
6β
-triol (1). 1 induced ALP activities of MC
3
T
3
-
E
1 cells and promoted cell proliferation. To investigate the relationships between the chemical structure and differentiation-inducing activity of the compound, ALP-inducing activities of MC
3
T
3
-
E
1 cells by 1, ergosterol (2), ergocalciferol (
3
), cholesta-
3β
,
5α
,
6β
-triol (4), 7-dehydrocholesterol (
5
) and cholecalciferol (
6
) were tested. The enzyme activities of MC
3
T
3
-
E
1 cells were increased
3
.0-fold by 10 μ
M 1 and 2.4-fold by 10 μ
M 4. However, 2,
3
,
5
and
6
did not induce MC
3
T
3
-
E
1 cell ALP activity at 0.1—10 μ
M. These results suggested that the OH groups at C-
5
and/or C-
6
of 1 and 4 played an important role in their differentiation-inducing activities on MC
3
T
3
-
E
1 cells. Furthermore, 1 suppressed induction of MC
3
T
3
-
E
1 cell apoptosis by serum starvation.
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