High-Density Microwell Chip for Culture and Analysis of Stem Cells
2009 (English)In: PLos ONE, ISSN 1932-6203, Vol. 4, no 9, e6997- p.Article in journal (Refereed) Published
With recent findings on the role of reprogramming factors on stem cells, in vitro screening assays for studying (de)differentiation is of great interest. We developed a miniaturized stem cell screening chip that is easily accessible and provides means of rapidly studying thousands of individual stem/progenitor cell samples, using low reagent volumes. For example, screening of 700,000 substances would take less than two days, using this platform combined with a conventional bio-imaging system. The microwell chip has standard slide format and consists of 672 wells in total. Each well holds 500 nl, a volume small enough to drastically decrease reagent costs but large enough to allow utilization of standard laboratory equipment. Results presented here include weeklong culturing and differentiation assays of mouse embryonic stem cells, mouse adult neural stem cells, and human embryonic stem cells. The possibility to either maintain the cells as stem/progenitor cells or to study cell differentiation of stem/progenitor cells over time is demonstrated. Clonality is critical for stem cell research, and was accomplished in the microwell chips by isolation and clonal analysis of single mouse embryonic stem cells using flow cytometric cell-sorting. Protocols for practical handling of the microwell chips are presented, describing a rapid and user-friendly method for the simultaneous study of thousands of stem cell cultures in small microwells. This microwell chip has high potential for a wide range of applications, for example directed differentiation assays and screening of reprogramming factors, opening up considerable opportunities in the stem cell field.
Place, publisher, year, edition, pages
2009. Vol. 4, no 9, e6997- p.
animal cell; article; biochip; cell cloning; cell culture; cell differentiation; cell screening; cell selection; controlled study; embryo; embryonic stem cell; female; flow cytometry; human; human cell; microwell chip; molecular imaging; mouse; neural stem cell; nonhuman; stem cell; animal; C57BL mouse; cell separation; culture technique; cytology; DNA microarray; equipment design; metabolism; methodology; nerve cell
IdentifiersURN: urn:nbn:se:kth:diva-11661DOI: 10.1371/journal.pone.0006997ISI: 000269796300013ScopusID: 2-s2.0-70349206057OAI: oai:DiVA.org:kth-11661DiVA: diva2:279093
QC 201007282009-12-012009-12-012011-10-31Bibliographically approved