AggreWell™

Recommended for:	For the formation of highly uniform sized embryoid bodies (EBs) from hPSCs and mPSCs.
Accessory Products:	• PBS (Catalog #37350) • DMEM/F-12 (Catalog #36254) • ACCUTASE™ (Catalog #07920) • Y-27632 (Catalog #07171/07172) • Trypan Blue (Catalog #07050) • 6-Well Ultra-Low Adherent Plates (Catalog #27145) • 37 µm Reversible Strainer, small and large (Catalog #27215/27250)
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Contains:	AggreWell™ plates with microwells
Equipment Required:	Low speed centrifuge (e.g. Beckman GS-6) with a swinging bucket rotor fitted with a plate holder
Legal Statement:	ACCUTASE™ is a registered trademark of Innovative Cell Technologies, Inc. San Diego, CA
Other cell culture media, reagents & supplies:	Cell cultureware & equipment
Application:	Differentiation
Area of Interest:	Embryonic stem cell & iPS cell research, Stem cell biology
Cell Type:	Embryonic stem cells & iPS cells
Popular Product Line:	AggreWell
Species:	Human, Mouse

Procedures and instruction manuals:

•  MANUAL: Reproducible and Unifrom Embryoid Bodies Using AggreWell™ Plates
•  PRODUCT INFORMATION SHEET - 27840, Lot# All:28784-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 27940, Lot# All:28784-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 27845, Lot# All:29832-PIS_1_1_0.pdf
•  PRODUCT INFORMATION SHEET - 27945, Lot# All:29832-PIS_1_1_0.pdf
•  PRODUCT INFORMATION SHEET - 27865, Lot# All:28508-PIS_1_1_0.pdf
•  PRODUCT INFORMATION SHEET - 27965, Lot# All:28508-PIS_1_1_0.pdf

Educational resources:

•  VIDEO: AggreWell™: Are You Making Embryoid Bodies?
•  WEBINAR: Standardizing hESC and hiPSC Maintenance and Differentiation
•  BROCHURE: Standardized Solutions for mESC and miPSC Research
•  BROCHURE: Standardized Solutions for hESC & hiPSC Research
•  BROCHURE: AggreWell™: Reproducible, Uniform Embryoid Bodies
•  SCIENTIFIC POSTER: AggreWell™400 generates a uniform population of embryoid bodies from human embryonic stem cells: Effects of modified mTeSR™ formulations on stability and growth of AggreWell™ generated embryoid bodies
•  WALLCHART: Pluripotent Stem Cell Biology Wallchart
•  TECH BULLETIN: Mouse embryonic stem cell differentiation into multiple tissue types (Catalog #29014)
•  MINI REVIEW: Mini-review: Induced pluripotent stem cells (Catalog #29143)
•  WEBINAR: AggreWell™ - A New Technology for Uniform Embryoid Bodies
•  SCIENTIFIC POSTER: Efficient production of neural progenitors from human pluripotent stem cells in AggreWell™400 and AggreWell™800 in a novel induction medium

This product has been used in:

1. Mark D Ungrin et al. Rational bioprocess design for human pluripotent stem cell expansion and endoderm differentiation based on cellular dynamics.Biotechnol Bioeng (December 2, 2011)
2. Erkan Kiris et al. Embryonic stem cell-derived motoneurons provide a highly sensitive cell culture model for botulinum neurotoxin studies, with implications for high-throughput drug discovery.Stem Cell Res 6 (3) 195-205 (May 2011)
3. Jakub Tolar et al. Induced pluripotent stem cells from individuals with recessive dystrophic epidermolysis bullosa.J Invest Dermatol 131 (4) 848-856 (April 2011)
4. Celine L Bauwens et al. Geometric Control of Cardiomyogenic Induction in Human Pluripotent Stem Cells.Tissue Eng Part A (April 25, 2011)
5. Brandoch D Cook et al. Smad1 signaling restricts hematopoietic potential after promoting hemangioblast commitment.Blood (April 22, 2011)
6. Ji-Eun Kim et al. Investigating synapse formation and function using human pluripotent stem cell-derived neurons.Proc Natl Acad Sci U S A 108 (7) 3005-3010 (February 15, 2011)
7. Andr??s M Bratt-Leal et al. Incorporation of biomaterials in multicellular aggregates modulates pluripotent stem cell differentiation.Biomaterials 32 (1) 48-56 (January 2011)
8. Masakazu Kamata et al. Live cell monitoring of hiPSC generation and differentiation using differential expression of endogenous microRNAs.PLoS One 5 (7) e11834 (2010)
9. Masakazu Kamata et al. Generation of human induced pluripotent stem cells bearing an anti-HIV transgene by a lentiviral vector carrying an internal murine leukemia virus promoter.Hum Gene Ther 21 (11) 1555-1567 (November 2010)
10. Michaela Kunova et al. Development of humanized culture medium with plant-derived serum replacement for human pluripotent stem cells.Reprod Biomed Online 21 (5) 676-686 (November 2010)
11. Jakub Tolar et al. Hematopoietic differentiation of induced pluripotent stem cells from patients with mucopolysaccharidosis type I (Hurler syndrome).Blood (October 29, 2010)
12. Thaddeus G Golos et al. Embryonic stem cells as models of trophoblast differentiation: progress, opportunities, and limitations.Reproduction 140 (1) 3-9 (July 1, 2010)
13. Franklin D West et al. Porcine Induced Pluripotent Stem Cells Produce Chimeric Offspring.Stem Cells Dev (April 9, 2010)
14. Saranna Brugger et al. The Seventh Annual Ion Channel Retreat Vancouver, Canada, June 29-July 1, 2009.Assay Drug Dev Technol 8 (1) 19-26 (February 2010)
15. Brandon D Markway et al. Enhanced Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells in Low Oxygen Environment Micropellet Cultures.Cell Transplant (October 29, 2009)
16. Knut Woltjen et al. piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells.Nature 458 (7239) 766-770 (March 1, 2009)

Background References:

1. Mark D Ungrin et al. Reproducible, ultra high-throughput formation of multicellular organization from single cell suspension-derived human embryonic stem cell aggregates.PLoS One 3 (2) e1565 (2008)

Product Name	Description	Catalog #
mTeSR®1	Defined, Feeder-Independent Maintenance Medium for hESCs and hiPSCs	05850
mFreSR®	Defined, Serum-Free Cryopreservation Medium for hESCs and hiPSCs	05854
TeSR™2	Animal Protein-Free, Defined, Feeder-independent Medium for Maintenance of Undifferentiated hESCs or hiPSCs.	05860
AggreWell™	Simple and Standardized Production of Embryoid Bodies	27845
Collagenase Type IV	Collagenase Type IV in DMEM/F-12 (1 mg/mL)	07909
Dispase (1 mg/mL)	Dispase 1 mg/mL	07923
0.1% Gelatin in Water	Coating for Cultureware	07903
MEM Non-Essential Amino Acid Solution (100X)	MEM Non-Essential Amino Acid Solution (100X)	07600
L-Glutamine	L-Glutamine (200 mM)	07100
Trypsin-EDTA (0.05%)	Trypsin-EDTA (0.05%)	07910
ES-Cult® Hematopoietic Differentiation Kit without Cytokines	For the In Vitro Differentiation of Mouse ESCs and iPSCs to Hematopoietic Cells	03160
Trypsin-EDTA (0.25%)	Trypsin-EDTA (0.25%)	07901
ES-Cult® Hematopoietic Differentiation Kit with Cytokines	For the In Vitro Differentiation of Mouse ESCs and iPSCs to Hematopoietic Cells	03161

Comparison of a conventional EB formation method with EB formation using AggreWell™

Comparison of a conventional EB formation method with EB formation using Aggrewell™.
A) EBs formed using conventional methods are heterogeneous in size and shape resulting in inefficient differentiation.
B) EBs formed using AggreWell™400 plates are uniform in size and shape.

EBs generated using AggreWell™ plates are able to differentiate into multiple cell types more efficiently

EBs generated using Aggrewell™ plates are able to differentiate into multiple cell types more efficiently.
A) Neural rosettes stained with Pax6 (green) and Sox2 (red).
B) Hematopoietic cells were detected using CFC assays.

EB size is tightly controlled using AggreWell™ plates

The size of EBs can easily be adjusted using Aggrewell™ plates. Here, EBs are formed by seeding the wells of the plate with varying concentrations of single cells The size of EBs can be tightly controlled using so that approximately
(A) 500 or (B) 2000 cells seed each microwell.
(C) The size of EBs can be tightly controlled using AggreWell plates, unlike scraping protocols which give a wider distribution.