Expression of Tight Junction Molecule In The Human Serum-Induced Aggregation of Human Abdominal Adipose-Derived Stem Cells In Vitro
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Previously we have shown that human abdominal adipose derived-stem cells (ADSCs) could aggregate duringthe high-density culture in the presence of human serum (HS). In the present study, we observed that human cord blood serum(CBS) and follicular fluid (HFF) also induced aggregation. Similarly, porcine serum could induce aggregation whereas bovineand sheep sera induced little aggregation. qRT-PCR analyses demonstrated that, compared to FBS-cultured ADSCs, HSculturedcells exhibited higher level of mRNA expression of CLDN3, -6, -7, -15, and -16 genes among the tight junctionproteins. ADSCs examined at the time of aggregation by culture with HS, BSA, HFF, CBS, or porcine serum showedsignificantly higher level of mRNA expression of JAM2 among JAM family members. In contrast, cells cultured in FBS,bovine serum or sheep serum, showed lower level of JAM2 expression. Immunocytochemical analyses demonstrated that theaggregates of HS-cultured cells (HS-Agg) showed intense staining against the anti-JAM2 antibody whereas neither nonaggregatedcells (HS-Ex) nor FBS-cultured cells exhibited weak staining. Western blot results showed that HS-Agg expressedJAM2 protein more prominently than HS-Ex and FBS-cultured cells, both of latter reveled weaker intensity. These resultssuggest that the aggregation property of ADSCs during high-density culture would be dependent on the specific componentsof serum, and that JAM2 molecule could play a role in the animal sera-induced aggregation in vitro.
Abstract
Previously we have shown that human abdominal adipose derived-stem cells (ADSCs) could aggregate duringthe high-density culture in the presence of human serum (HS). In the present study, we observed that human cord blood serum(CBS) and follicular fluid (HFF) also induced aggregation. Similarly, porcine serum could induce aggregation whereas bovineand sheep sera induced little aggregation. qRT-PCR analyses demonstrated that, compared to FBS-cultured ADSCs, HSculturedcells exhibited higher level of mRNA expression of CLDN3, -6, -7, -15, and -16 genes among the tight junctionproteins. ADSCs examined at the time of aggregation by culture with HS, BSA, HFF, CBS, or porcine serum showedsignificantly higher level of mRNA expression of JAM2 among JAM family members. In contrast, cells cultured in FBS,bovine serum or sheep serum, showed lower level of JAM2 expression. Immunocytochemical analyses demonstrated that theaggregates of HS-cultured cells (HS-Agg) showed intense staining against the anti-JAM2 antibody whereas neither nonaggregatedcells (HS-Ex) nor FBS-cultured cells exhibited weak staining. Western blot results showed that HS-Agg expressedJAM2 protein more prominently than HS-Ex and FBS-cultured cells, both of latter reveled weaker intensity. These resultssuggest that the aggregation property of ADSCs during high-density culture would be dependent on the specific componentsof serum, and that JAM2 molecule could play a role in the animal sera-induced aggregation in vitro.
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