MesenCult®-XF Culture Kit

Recommended for:	• Mesenchymal stem cell expansion from primary human bone marrow • Long-term culture of mesenchymal stem cells (greater than 8 passages) • Detection and enumeration of CFU-F from primary human bone marrow and culture-expanded mesenchymal stem cells
Components:	• MesenCult®-XF Basal Medium, 400 mL (Catalog #05421) • MesenCult®-XF Supplement (5X), 100 mL (Catalog #05422) • MesenCult®-XF Attachment Substrate, 5 mg (Catalog #05424)
Accessory Products:	• MesenCult®-ACF Dissociation Kit (Catalog #05426) • L-glutamine (Catalog #07100)
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Product Type:	Specialized cell culture media
Application:	Colony assays & quantification, Expansion
Area of Interest:	Mesenchymal stem cell research, Stem cell biology
Cell Type:	Mesenchymal stem cells
Medium Type:	Serum-free, Liquid
Popular Product Line:	MesenCult
Species:	Human

Procedures and instruction manuals:

•  MANUAL: Enumeration and Expansion of Human Mesenchymal Stem Cells Using MesenCult®-XF Medium
•  PRODUCT INFORMATION SHEET - 05429, Lot# All:29919-PIS_2_1_1.pdf
•  PRODUCT INFORMATION SHEET - 05429, Lot# All:28419-PIS_1_0_1.pdf
•  PRODUCT INFORMATION SHEET - 05429, Lot# All:29921-PIS_1_2_0.pdf

Educational resources:

•  VIDEO: MesenCult®-XF Introduction: A Defined, Xeno-free Medium for Human MSCs
•  BROCHURE: Mesenchymal Stem Cells: Standardized Media and Reagents Erratum: Figures 6C and 6F on page 7 of the brochure should be labeled as "Passage 8", not "Passage 6".
•  BROCHURE: MesenCult™-XF: Defined, Xeno-Free Culture of Human Mesenchymal Stem Cells
•  SCIENTIFIC POSTER: Expansion of mesenchymal progenitor cells from human bone marrow in a novel serum- and animal component-free culture medium
•  SCIENTIFIC POSTER: Characterization of human bone marrow-derived mesenchymal progenitor cells in a novel serum- and animal component-free culture medium
•  SCIENTIFIC POSTER: Expansion and immunosuppressive ability of mesenchymal progenitor cells from human bone marrow cultured in novel serum and animal component-free culture medium
•  MINI REVIEW: Mini-review: Mesenchymal cells (Catalog #29018)
•  WEBINAR: Characterization of MSCs Cultured in Xeno-Free MesenCult®-XF
•  TECH BULLETIN: Characterization of Human MSCs Cultured in MesenCult®-XF (Catalog #28308)
•  WALLCHART: The Identity and Properties of Mesenchymal Stem Cells Wallchart

MSDS:

•  05422-MSDS.pdf
•  05421-MSDS.pdf
•  05425-MSDS.pdf

This product has been used in:

1. Hartmann I et al. Umbilical cord tissue-derived mesenchymal stem cells grow best under GMP-compliant culture conditions and maintain their phenotypic and functional propertiesJ Immunol Methods (October 27, 2010)

Background References:

1. M Dominici et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.Cytotherapy 8 (4) 315-317 (2006)

Product Name	Description	Catalog #
MesenCult®-XF Medium	Defined, Xeno-Free Medium for Human Mesenchymal Stem Cells	05420
MesenCult®-XF Attachment Substrate	Xeno-Free Attachment Substrate for Human Mesenchymal Stem Cells	05424
MesenCult®-ACF Dissociation Kit	Dissociation Kit for Human Mesenchymal Stem Cells	05426
MesenCult® Proliferation Kit (Human)	Medium for Detection of CFU-F and Expansion of Human Mesenchymal Stem Cells	05411
MesenCult® Adipogenic Stimulatory Supplements (Human)	Supplement for Differentiating Human Mesenchymal Stem Cells into Adipocytes	05403
EasySep® Human CD271 Selection Kit	Immunomagnetic Positive Selection Kit	18659
L-Glutamine	L-Glutamine (200 mM)	07100

Human bone marrow-derived MSCs cultured in MesenCult®-XF expand faster than cells cultured in a traditional serum-based medium (n = 3; Mean ± SEM)

Human bone marrow-derived MSCs cultured in MesenCult®-XF display multi-lineage differentiation potential

A. Oil red O staining of adipocytes generated from passage 1 MSCs previously cultured in MesenCult®-XF.B. Alizarin Red detection of Ca2+ deposits indicates the formation of bone structures in cells generated from passage 4 MSCs previously cultured in MesenCult®-XF.C. Alcian Blue staining of chondrocytes generated from passage 2 MSCs previously cultured in MesenCult®-XF.D. Collagen II staining of chondrocytes generated from passage 2 MSCs previously cultured in MesenCult®-XF.

Primary human bone marrow-derived MSCs show less hematopoietic cell contamination when cultured in (A) MesenCult®-XF compared to a (B) traditional serum-based medium

Passage 0 human bone marrow-derived MSCs cultured in MesenCult®-XF (A) or a traditional serum-based medium (B).

Phenotype of culture-expanded human MSCs cultured in MesenCult®-XF.

MSCs cultured in MesenCult®-XF suppress T cell proliferation and reduce cell cycle division more robustly than MSCs cultured in a traditional serum-based medium

Passage 2 MSCs generated in MesenCult®-XF or a traditional serum-based medium were treated with mitomycin C prior to co-culture with T cells. T cells were purified from peripheral blood using EasySep® (Catalog #19051) immunomagnetic separation and fluorescently labeled using 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester (5(6)CFDA,SE; CFSE). 2 x 10e5 CFSE-labeled T cells were cultured with 1 x 10e5 MSCs in serum-free medium supplemented with 100 U/mL IL-2. T cells were stimulated with tetrameric antibody complexes against CD3ε, CD28 and CD2. On days 3 and 7, cells were harvested, stained with anti-CD45 antibody and propidium iodide and the T cell division history measured as CFSE dye dilution analyzed by flow cytometry.