Background: Phospholipase A2 (PLA2) is involved in inflammation and cell death following stroke, ... more Background: Phospholipase A2 (PLA2) is involved in inflammation and cell death following stroke, and inhibition of its activity may promote neuroregeneration. This study aimed to observe the influence of Acalypha indica Linn root extract towards relative cell viability and PLA2 enzyme level in post-hypoxic hippocampal tissue culture. Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from Sprague Dawley rat exposed to hypoxia with 5% O2 / 5% CO2 / N2 balanced gas for 24 hours. Post-hypoxia, Acalypha indica Linn root extract was added at doses of 10, 15, and 20 mg/mL to three treatment groups. No treatment was given to the control group. Each group consists of six samples. After 72 hours of incubation, relative cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and phospholipase A2 enzyme level was determined using ELISA. Results: PLA2 enzyme level of rat hippocampal tissue culture treated with Acalypha indica Linn root extract at 10, 15, and 20 mg/mL were significantly lower than that of control (5.55 ng/mL, 6.85 ng/mL, and 7.42 ng/mL vs. 7.96 ng/mL, p < 0.05). Conclusion: Acalypha indica Linn root extract increases the relative cell viability and decreases the PLA2 enzyme level of post-hypoxic mouse hippocampal tissue with the optimal dose of the extract at 10 mg/mL
BACKGROUND Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are ea... more BACKGROUND Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC). METHODS In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4) expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1) proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. RESULTS Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05). Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity CONCLUSIONS Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management. Key words: Fibroblast, PBMC, co-culture, foreskin
Background: Phospholipase A2 (PLA2) is involved in inflammation and cell death following stroke, ... more Background: Phospholipase A2 (PLA2) is involved in inflammation and cell death following stroke, and inhibition of its activity may promote neuroregeneration. This study aimed to observe the influence of Acalypha indica Linn root extract towards relative cell viability and PLA2 enzyme level in post-hypoxic hippocampal tissue culture. Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from Sprague Dawley rat exposed to hypoxia with 5% O2 / 5% CO2 / N2 balanced gas for 24 hours. Post-hypoxia, Acalypha indica Linn root extract was added at doses of 10, 15, and 20 mg/mL to three treatment groups. No treatment was given to the control group. Each group consists of six samples. After 72 hours of incubation, relative cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and phospholipase A2 enzyme level was determined using ELISA. Results: PLA2 enzyme level of rat hippocampal tissue culture treated with Acalypha indica Linn root extract at 10, 15, and 20 mg/mL were significantly lower than that of control (5.55 ng/mL, 6.85 ng/mL, and 7.42 ng/mL vs. 7.96 ng/mL, p < 0.05). Conclusion: Acalypha indica Linn root extract increases the relative cell viability and decreases the PLA2 enzyme level of post-hypoxic mouse hippocampal tissue with the optimal dose of the extract at 10 mg/mL
BACKGROUND Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are ea... more BACKGROUND Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC). METHODS In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4) expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1) proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. RESULTS Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05). Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity CONCLUSIONS Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management. Key words: Fibroblast, PBMC, co-culture, foreskin
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Papers by Indra Kusuma
activity may promote neuroregeneration. This study aimed to observe the influence of Acalypha indica Linn root extract
towards relative cell viability and PLA2 enzyme level in post-hypoxic hippocampal tissue culture.
Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from Sprague Dawley rat
exposed to hypoxia with 5% O2 / 5% CO2 / N2 balanced gas for 24 hours. Post-hypoxia, Acalypha indica Linn root
extract was added at doses of 10, 15, and 20 mg/mL to three treatment groups. No treatment was given to the control
group. Each group consists of six samples. After 72 hours of incubation, relative cell viability was measured using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and phospholipase A2 enzyme level
was determined using ELISA.
Results: PLA2 enzyme level of rat hippocampal tissue culture treated with Acalypha indica Linn root extract at 10, 15,
and 20 mg/mL were significantly lower than that of control (5.55 ng/mL, 6.85 ng/mL, and 7.42 ng/mL vs. 7.96 ng/mL,
p < 0.05).
Conclusion: Acalypha indica Linn root extract increases the relative cell viability and decreases the PLA2 enzyme
level of post-hypoxic mouse hippocampal tissue with the optimal dose of the extract at 10 mg/mL
METHODS In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4) expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1) proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test.
RESULTS Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05). Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity
CONCLUSIONS Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.
Key words: Fibroblast, PBMC, co-culture, foreskin
activity may promote neuroregeneration. This study aimed to observe the influence of Acalypha indica Linn root extract
towards relative cell viability and PLA2 enzyme level in post-hypoxic hippocampal tissue culture.
Methods: Experimental in vitro study using 24 primary neuronal cell cultures obtained from Sprague Dawley rat
exposed to hypoxia with 5% O2 / 5% CO2 / N2 balanced gas for 24 hours. Post-hypoxia, Acalypha indica Linn root
extract was added at doses of 10, 15, and 20 mg/mL to three treatment groups. No treatment was given to the control
group. Each group consists of six samples. After 72 hours of incubation, relative cell viability was measured using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) examination, and phospholipase A2 enzyme level
was determined using ELISA.
Results: PLA2 enzyme level of rat hippocampal tissue culture treated with Acalypha indica Linn root extract at 10, 15,
and 20 mg/mL were significantly lower than that of control (5.55 ng/mL, 6.85 ng/mL, and 7.42 ng/mL vs. 7.96 ng/mL,
p < 0.05).
Conclusion: Acalypha indica Linn root extract increases the relative cell viability and decreases the PLA2 enzyme
level of post-hypoxic mouse hippocampal tissue with the optimal dose of the extract at 10 mg/mL
METHODS In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4) expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1) proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test.
RESULTS Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05). Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity
CONCLUSIONS Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.
Key words: Fibroblast, PBMC, co-culture, foreskin