StemXVivo Hepatocyte Differentiation Kit

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME. 

A: The cells are differentiated to the definitive endoderm stage by around day 4 from differentiation initiation. A schematic of the differentiation progression can be viewed in this scientific poster: https://resources.rndsystems.com/images/site/rnd-systems-stem-cell-derived-hepatocyte-like-cells-isscr-2016.pdf

A:   Yes. Our labs have used iPSC cells to differentiate into hepatocytes with this kit without significant modification of the protocol provided in the SC033 kit booklet.  Plating density would be one parameter to optimize, since growth rate and cell size can differ between cell lines. Ideally, sufficient cells should be plated so that 80-90% confluency is reached 24-48 hrs after plating.   Also, there is some variability in how well different iPSC lines differentiate into different lineages.
 

A: The differentiated (Day 19) hepatocytes can be harvested using a 1:1 mixture of TrypLE Express and Accutase for 5 minutes at 37°C.

A: For re-plating the day 19 hepatocytes, we used plates coated with Ultimatrix (catalog # BME001) or other Cutltrex BME. The medium used was Differentiation Base media II + Differentiation Cocktail IV (as described in Catalog # SC033).  The cell density ranged from 3 x104 to  2 x 105. Addition of dexamethasone (50 µM) improved albumin expression.

A: We have been successful in re-plating the Day 19 hepatocytes cells and maintaining the differentiated hepatocytes for 10 additional days.

A: In-house, we have been able to cryopreserve day 9 progenitors. 

A: We have not done any gene editing work in-house but citations for CRISPR hepatocytes are available. https://www.sciencedirect.com/science/article/pii/S2589555921001658

A: We have not needed to adapt iPSCs prior to differentiation, however, if iPSCs have been grown with Matrigel it may be helpful to have 1-2 passages in Cultrex BME.