Human bone marrow-derived mesenchymal stem cell gene expression patterns vary with culture conditions.
- Author:
Myoung Woo LEE
1
;
Dae Seong KIM
;
Keon Hee YOO
;
Hye Ryung KIM
;
In Keun JANG
;
Ji Hyang LEE
;
So Yeon KIM
;
Meong Hi SON
;
Soo Hyun LEE
;
Hye Lim JUNG
;
Ki Woong SUNG
;
Hong Hoe KOO
Author Information
- Publication Type:Original Article
- Keywords: Mesenchymal stem cell; Gene expression pattern; Seeding density; Culture time; Cell therapy
- MeSH: Cell Count; Cell Movement; Cyclooxygenase 1; Gene Expression; Hepatocyte Growth Factor; Humans; Immunomodulation; Indoleamine-Pyrrole 2,3,-Dioxygenase; Interferon-gamma; Leukemia Inhibitory Factor; Mesenchymal Stromal Cells; Polymerase Chain Reaction; Population Characteristics; Regeneration; RNA, Messenger; Seeds; Tissue Therapy
- From:Blood Research 2013;48(2):107-114
- CountryRepublic of Korea
- Language:English
- Abstract: BACKGROUND: Because of the heterogeneity of human mesenchymal stem cells (MSCs), methods for cell expansion in culture and the effects on gene expression are critical factors that need to be standardized for preparing MSCs. We investigated gene expression patterns of MSCs with different seeding densities and culture times. METHODS: Bone marrow-derived MSCs were plated at densities from 200 cells/cm2 to 5,000 cells/cm2, and the gene expression patterns were evaluated over time using a reverse-transcription polymerase chain reaction assay. RESULTS: The mRNA levels of factors that play a critical role in cell migration and tissue regeneration, such as podocalyxin-like protein (PODXL), alpha4-integrin, alpha6-integrin, and leukemia inhibitory factor (LIF), were higher in MSCs plated at 200 cells/cm2 than in MSCs plated at 5,000 cells/cm2. The mRNA levels of these factors gradually increased for 10 days and then decreased by day 15 in culture. MSCs seeded at 200 cells/cm2 that were cultured for 10 days expressed high levels of Oct-4 and Nanog. Indoleamine 2,3-dioxygenase, cyclooxygenase-1, and hepatocyte growth factor expression were upregulated in the presence of the proinflammatory cytokine interferon-gamma in these cells. CONCLUSION: We found differences in the gene expression patterns of MSCs under different culture conditions. MSCs from 10-day cultures seeded at a low density were efficiently expanded, expressed PODXL, alpha6-integrin, alpha4-integrin, and LIF, and maintained properties like stemness and immunomodulation. Therefore, ex vivo expansion of MSCs maintained for an adequate culture time after plating at low cell density can provide an effective regenerative medicinal strategy for cell therapies using MSCs.
