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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

Difference of gene expressions in four different BM cell populations in human isletsIsolated RNA from human islet tissue after culture 4 weeks was evaluated for transcriptional factors activation by RT-PCR. The changes show to comparison to islet only group. Only two factor activations were detected in EDCs group (A), three in MSCs group (B) and six in BM group (C). (* = p<0.05 n = 3)
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Figure 4: Difference of gene expressions in four different BM cell populations in human isletsIsolated RNA from human islet tissue after culture 4 weeks was evaluated for transcriptional factors activation by RT-PCR. The changes show to comparison to islet only group. Only two factor activations were detected in EDCs group (A), three in MSCs group (B) and six in BM group (C). (* = p<0.05 n = 3)

Mentions: A total of 24 β-cell differentiation and insulin related genes were evaluated and analyzed on culture day 28. The results showed that mwBM co-cultured islets induced the activation of six factors: Isl1, MAFA, Nkx6.1, PAX6, IPF1 and down-regulated Kif4a vs. islet only culture (p<0.05, Fig. 4). However, E cocultured islet induced two factor expressions Glucagon (GCG) and the paired box family factors 4 (PAX4) vs. islet only culture. M cocultured islet induced three positive activations: islet amyloid polypeptide (IAPP), PAX4 and the NK-homeodomain factor 2 (NKx.2) vs. islet only culture (p< 0.05). (Reference to these factors: Pandian GN, Taniguchi J, Sugiyama H. Cellular reprogramming for pancreatic β-cell regeneration: clinical potential of small moleculae control. Clin Tansl Medicine 2014, 3:6).


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)

Difference of gene expressions in four different BM cell populations in human isletsIsolated RNA from human islet tissue after culture 4 weeks was evaluated for transcriptional factors activation by RT-PCR. The changes show to comparison to islet only group. Only two factor activations were detected in EDCs group (A), three in MSCs group (B) and six in BM group (C). (* = p<0.05 n = 3)
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Difference of gene expressions in four different BM cell populations in human isletsIsolated RNA from human islet tissue after culture 4 weeks was evaluated for transcriptional factors activation by RT-PCR. The changes show to comparison to islet only group. Only two factor activations were detected in EDCs group (A), three in MSCs group (B) and six in BM group (C). (* = p<0.05 n = 3)
Mentions: A total of 24 β-cell differentiation and insulin related genes were evaluated and analyzed on culture day 28. The results showed that mwBM co-cultured islets induced the activation of six factors: Isl1, MAFA, Nkx6.1, PAX6, IPF1 and down-regulated Kif4a vs. islet only culture (p<0.05, Fig. 4). However, E cocultured islet induced two factor expressions Glucagon (GCG) and the paired box family factors 4 (PAX4) vs. islet only culture. M cocultured islet induced three positive activations: islet amyloid polypeptide (IAPP), PAX4 and the NK-homeodomain factor 2 (NKx.2) vs. islet only culture (p< 0.05). (Reference to these factors: Pandian GN, Taniguchi J, Sugiyama H. Cellular reprogramming for pancreatic β-cell regeneration: clinical potential of small moleculae control. Clin Tansl Medicine 2014, 3:6).

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