Two steps to functional mesenchymal stromal cells for clinical application

Christina Bartmann, Eva Rohde, Katharina Schallmoser, Peter Pürstner, Gerhard Lanzer, Werner Linkesch, Dirk Strunk
Transfusion 2007, 47 (8): 1426-35

BACKGROUND: Ex vivo expansion of multipotent mesenchymal stromal cells (MSCs) is a prerequisite for evaluating their therapeutic potential in ongoing clinical trials. Even large volumes of starting material and extended culture periods, however, do not necessarily produce 2 x 10(6) MSCs per kg per adult patient. A new two-step procedure has been devised to propagate more than 1 x 10(8) MSCs from small marrow volumes within fewer than 4 weeks.

STUDY DESIGN AND METHODS: The influence of log fold decreased MSC seeding (2500, 250, 25, 2.5/cm(2)) on clinical-scale expansion, MSC phenotype, and immunomodulatory function combined with multiplex cytokine display was analyzed. Maintenance of MSC characteristics was tested in fibroblast colony-forming unit and differentiation assays.

RESULTS: Reduced seeding density boosted MSC propagation. Low-density expanded MSCs were CD29+, CD73+, CD90+, CD105+, CD14-, CD34-, CD45-, HLA-DR-; retained their differentiation potential; and inhibited lymphocyte proliferation. This was accompanied by deregulated cytokine production. Seeding 0.7 x 10(6) to 1 x 10(6) MSCs derived from a 10- to 13-day primary culture at a low density of 28 to 40 per cm(2) permitted propagation of 1.5 x 10(8) to 3.7 x 10(8) functional MSCs within a 13- to 15-day secondary expansion step.

CONCLUSION: Primary seeding of only 10-mL marrow aspirates on approximately 0.2-m(2) culture area (Step 1) followed by expansion on 2.5 m(2) (Step 2) is sufficient to consistently generate at least 1.5 x 10(8) MSCs in fetal bovine serum-supplemented medium within less than 4 weeks. The efficiency of this two-step procedure for clinical-scale MSC propagation may facilitate rational clinical testing of MSC-based therapies.

Full Text Links

Find Full Text Links for this Article


You are not logged in. Sign Up or Log In to join the discussion.

Related Papers

Available on the App Store

Available on the Play Store
Remove bar
Read by QxMD icon Read

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"