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Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro.

Luo LG, Xiong F, Ravassard P, Luo JZ - Br J Med Med Res (2015)

Bottom Line: However, only whole BM enables to sustain the capability of islet β-cell self regeneration resulting in increasing β cell population while single E and M individual do not significantly affect on that.Mechanism approach to explore β-cell self regeneration by evaluating transcription factor expressions, we found that BM significantly increases the activations of β-cell regeneration relative transcription factors, the LIM homeodomain protein (Isl1), homologue to zebrafish somite MAF1 (MAFa), the NK-homeodomain factor 6.1 (NKX6.1), the paired box family factors 6 (PAX6), insulin promoter factor 1 (IPF1) and kinesin family member 4A (KIF4a).These results suggest that BM and its derived M and E cells enable to support human islet β-cell function.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine/Research, Roger Williams Medical Center, Boston University, USA.

ABSTRACT

Aims: Allogeneic bone marrow (BM) has been shown to support human islet survival and function in long-term culture by initiating human islet vascularization and β-cell regeneration. Various BM subpopulations may play different roles in human islet functions and survival. In this paper we investigated the effects of BM and its subpopulations, endothelial progenitor cells (E) and mesenchymal (M) cells on human islet's β-cell function and regeneration.

Study design: Isolation and identification of subpopulations from human bone marrow and culture with allogeneic human islet to investigate effects of different cell population on human islet function and regeneration.

Place and duration of study: Department of Medicine, Center for Stem Cell & Diabetes Research, RWMC, Providence, RI, USA, between 2010 - 2014.

Methodology: Human islets were distributed from Integrated Islet Distribution Program (IIDP) and human bone marrow (BM) was harvested by Bone marrow transplantation center at Roger Williams Hospital. BM subpopulation was identified cell surface markers through Fluorescence-activated cell sorting, applied in flow cytometry (FACS), islet function was evaluated by human ELISA kit and β cell regeneration was evaluated by three methods of Cre-Loxp cell tracing, β cell sorting and RT-PCR for gene expression.

Results: Four different BM and seven different islet donates contributed human tissues. We observed islet β-cell having self regeneration capability in short term culture (3∼5 days) using a Cre-Loxp cell tracing. BM and its subtype E, M have similar benefits on β cell function during co-culture with human islet comparison to islet only. However, only whole BM enables to sustain the capability of islet β-cell self regeneration resulting in increasing β cell population while single E and M individual do not significantly affect on that. Mechanism approach to explore β-cell self regeneration by evaluating transcription factor expressions, we found that BM significantly increases the activations of β-cell regeneration relative transcription factors, the LIM homeodomain protein (Isl1), homologue to zebrafish somite MAF1 (MAFa), the NK-homeodomain factor 6.1 (NKX6.1), the paired box family factors 6 (PAX6), insulin promoter factor 1 (IPF1) and kinesin family member 4A (KIF4a).

Conclusion: These results suggest that BM and its derived M and E cells enable to support human islet β-cell function. However, only BM can sustain the capability of β-cell self regeneration through initiating β-cell transcriptional factors but not individual E and M cells suggesting pure E and M cells less supportive for islet long-term survival in vitro.

No MeSH data available.


Related in: MedlinePlus

Evaluation effects of E, M and mwBM on human islet β cell functionAfter 4 weeks culture, human islet β-cell function was evaluated by human insulin ELISA assay. the results indicated that all three cell populations in culture promotes human islet β-cell function vs. islet only culture 1637.42 μU/ml) (P<0.01, N = 3). however, no significant difference of M, E was found along with mwBM groups (6526.01, 7900.25 and 7882.47μU/ml)
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Figure 2: Evaluation effects of E, M and mwBM on human islet β cell functionAfter 4 weeks culture, human islet β-cell function was evaluated by human insulin ELISA assay. the results indicated that all three cell populations in culture promotes human islet β-cell function vs. islet only culture 1637.42 μU/ml) (P<0.01, N = 3). however, no significant difference of M, E was found along with mwBM groups (6526.01, 7900.25 and 7882.47μU/ml)

Mentions: Using the same culture protocol for BM/human islet co-culture, three different subpopulations, mwBM, purified M and E, from same bone marrow donor were co-cultured with human islets from same donor. Co-cultured tissue medium was collected on day 30. Human islet insulin release in medium was assayed by human insulin kit and results show that co-culture averaged insulin release no statistically significant difference amongst the three groups: mwBM (7882.47±1647.17 μU/ml), E (7900.25±1149.27 μU/ml) and M (6526.01±542.42 μU/ml). However, all three groups showed statistically elevated levels of insulin when compared to the islet only group (1637.42±33.35 μU/ml) p< 0.01 n = 18. (Fig. 2).


Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro.

Luo LG, Xiong F, Ravassard P, Luo JZ - Br J Med Med Res (2015)

Evaluation effects of E, M and mwBM on human islet β cell functionAfter 4 weeks culture, human islet β-cell function was evaluated by human insulin ELISA assay. the results indicated that all three cell populations in culture promotes human islet β-cell function vs. islet only culture 1637.42 μU/ml) (P<0.01, N = 3). however, no significant difference of M, E was found along with mwBM groups (6526.01, 7900.25 and 7882.47μU/ml)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4837454&req=5

Figure 2: Evaluation effects of E, M and mwBM on human islet β cell functionAfter 4 weeks culture, human islet β-cell function was evaluated by human insulin ELISA assay. the results indicated that all three cell populations in culture promotes human islet β-cell function vs. islet only culture 1637.42 μU/ml) (P<0.01, N = 3). however, no significant difference of M, E was found along with mwBM groups (6526.01, 7900.25 and 7882.47μU/ml)
Mentions: Using the same culture protocol for BM/human islet co-culture, three different subpopulations, mwBM, purified M and E, from same bone marrow donor were co-cultured with human islets from same donor. Co-cultured tissue medium was collected on day 30. Human islet insulin release in medium was assayed by human insulin kit and results show that co-culture averaged insulin release no statistically significant difference amongst the three groups: mwBM (7882.47±1647.17 μU/ml), E (7900.25±1149.27 μU/ml) and M (6526.01±542.42 μU/ml). However, all three groups showed statistically elevated levels of insulin when compared to the islet only group (1637.42±33.35 μU/ml) p< 0.01 n = 18. (Fig. 2).

Bottom Line: However, only whole BM enables to sustain the capability of islet β-cell self regeneration resulting in increasing β cell population while single E and M individual do not significantly affect on that.Mechanism approach to explore β-cell self regeneration by evaluating transcription factor expressions, we found that BM significantly increases the activations of β-cell regeneration relative transcription factors, the LIM homeodomain protein (Isl1), homologue to zebrafish somite MAF1 (MAFa), the NK-homeodomain factor 6.1 (NKX6.1), the paired box family factors 6 (PAX6), insulin promoter factor 1 (IPF1) and kinesin family member 4A (KIF4a).These results suggest that BM and its derived M and E cells enable to support human islet β-cell function.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine/Research, Roger Williams Medical Center, Boston University, USA.

ABSTRACT

Aims: Allogeneic bone marrow (BM) has been shown to support human islet survival and function in long-term culture by initiating human islet vascularization and β-cell regeneration. Various BM subpopulations may play different roles in human islet functions and survival. In this paper we investigated the effects of BM and its subpopulations, endothelial progenitor cells (E) and mesenchymal (M) cells on human islet's β-cell function and regeneration.

Study design: Isolation and identification of subpopulations from human bone marrow and culture with allogeneic human islet to investigate effects of different cell population on human islet function and regeneration.

Place and duration of study: Department of Medicine, Center for Stem Cell & Diabetes Research, RWMC, Providence, RI, USA, between 2010 - 2014.

Methodology: Human islets were distributed from Integrated Islet Distribution Program (IIDP) and human bone marrow (BM) was harvested by Bone marrow transplantation center at Roger Williams Hospital. BM subpopulation was identified cell surface markers through Fluorescence-activated cell sorting, applied in flow cytometry (FACS), islet function was evaluated by human ELISA kit and β cell regeneration was evaluated by three methods of Cre-Loxp cell tracing, β cell sorting and RT-PCR for gene expression.

Results: Four different BM and seven different islet donates contributed human tissues. We observed islet β-cell having self regeneration capability in short term culture (3∼5 days) using a Cre-Loxp cell tracing. BM and its subtype E, M have similar benefits on β cell function during co-culture with human islet comparison to islet only. However, only whole BM enables to sustain the capability of islet β-cell self regeneration resulting in increasing β cell population while single E and M individual do not significantly affect on that. Mechanism approach to explore β-cell self regeneration by evaluating transcription factor expressions, we found that BM significantly increases the activations of β-cell regeneration relative transcription factors, the LIM homeodomain protein (Isl1), homologue to zebrafish somite MAF1 (MAFa), the NK-homeodomain factor 6.1 (NKX6.1), the paired box family factors 6 (PAX6), insulin promoter factor 1 (IPF1) and kinesin family member 4A (KIF4a).

Conclusion: These results suggest that BM and its derived M and E cells enable to support human islet β-cell function. However, only BM can sustain the capability of β-cell self regeneration through initiating β-cell transcriptional factors but not individual E and M cells suggesting pure E and M cells less supportive for islet long-term survival in vitro.

No MeSH data available.


Related in: MedlinePlus