Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

Ren, Hongzheng; Cao, Ying; Zhao, Qinjun; Li, Jing; Zhou, Chenyu; Liao, Lianming; Jia, Mingyue; Zhao, Qian; Cai, Huiguo; Han, Zhongchao; Yang, Renchi; Chen, Guoqiang; Zhao, Robert Chunhua

doi:10.1016/j.bbrc.2006.05.169

Cited by 184 publications

(165 citation statements)

References 33 publications

Supporting

Mentioning

142

Contrasting

Unclassified

Order By: Relevance

“…We demonstrated here that BM-MSCs cultured under hypoxia had higher proliferation rates than BM-MSCs cultured under normoxia. This finding is consistent with previous reports that demonstrated increased proliferation of BM-MSCs cultured under hypoxia (Ren et al 2006;Grayson et al 2007). In stem cells, maintenance of proliferation is affected by molecular mechanisms mediated by stemness genes, such as KLF4, OCT4, and C-MYC.…”

Section: Discussionsupporting

confidence: 93%

Effect of low oxygen tension on the biological characteristics of human bone marrow mesenchymal stem cells

Kim

Lee

et al. 2016

Cell Stress and Chaperones

View full text Add to dashboard Cite

Culture of mesenchymal stem cells (MSCs) under ambient conditions does not replicate the low oxygen environment of normal physiological or pathological states and can result in cellular impairment during culture. To overcome these limitations, we explored the effect of hypoxia (1 % O 2 ) on the biological characteristics of MSCs over the course of different culture periods. The following biological characteristics were examined in human bone marrow-derived MSCs cultured under hypoxia for 8 weeks: proliferation rate, morphology, cell size, senescence, immunophenotypic characteristics, and the expression levels of stemness-associated factors and cytokine and chemokine genes. MSCs cultured under hypoxia for approximately 2 weeks showed increased proliferation and viability. During long-term culture, hypoxia delayed phenotypic changes in MSCs, such as increased cell volume, altered morphology, and the expression of senescence-associated-β-gal, without altering their characteristic immunophenotypic characteristics. Furthermore, hypoxia increased the expression of stemness and chemokine-related genes, including OCT4 and CXCR7, and did not decrease the expression of KLF4, C-MYC, CCL2, CXCL9, CXCL10, and CXCR4 compared with levels in cells cultured under normoxia. In conclusion, low oxygen tension improved the biological characteristics of MSCs during ex vivo expansion. These data suggest that hypoxic culture could be a useful method for increasing the efficacy of MSC cell therapies.

show abstract

Section: Discussionsupporting

confidence: 93%

Effect of low oxygen tension on the biological characteristics of human bone marrow mesenchymal stem cells

Kim

Lee

et al. 2016

Cell Stress and Chaperones

View full text Add to dashboard Cite

show abstract

“…During in vitro culture, hDSPCs, which exhibited stem celllike characteristics in our previous study, gradually lose the expression of transcription factors associated with multipotency and self-renewal, such as SOX2 and NANOG for maintaining the characteristics of stem cells. Several previous reports suggested that the quality of the stem cells strongly depend on the culture method (Driskell et al, 2012;Grayson et al, 2007;Kuroda et al, 2010;Minchenko et al, 1994;Reiter et al, 2008;Ren et al, 2006;Rendl et al, 2005). Thus, we investigated whether hDSPCs retain their stemness in various culture conditions despite long-term culture.…”

Section: Discussionmentioning

confidence: 92%

“…It regulates various cellular events critical to the in vitro growth of cells. In the previous studies, hypoxia enhances the stemness of bone marrow mesenchymal stem cells (Grayson et al, 2007;Ren et al, 2006). Thus, we examined whether O 2 concentration had effects on preserving the stemness of hDSPCs.…”

Section: Discussionmentioning

confidence: 94%

“…Recent reports suggested that hypoxia contributes to maintain the stemness of MSCs (Grayson et al, 2007;Ren et al, 2006). Thus, we tested the effects of oxygen level on dermal progenitor markers and found that there were no significant differences between normoxic (20% of O 2 ) and hypoxic (1% or 5% of O 2 ) conditions in the mRNA expression levels of SOX2, NANOG and S100B (Supplementary Figs.…”

Section: Effects Of Oxygen In Maintaining the Stemness Of Hdspcsmentioning

confidence: 99%

See 1 more Smart Citation

Novel In Vitro Culture Condition Improves the Stemness of Human Dermal Stem/Progenitor Cells

Shim

Lee

Shin

2013

Molecules and Cells

View full text Add to dashboard Cite

Cell therapy using adult stem cells has emerged as a potentially new approach for the treatment of various diseases. Therefore, it is an essential procedure to maintain the stemness of adult stem cells for clinical treatment. We previously reported that human dermal stem/progenitor cells (hDSPCs) can be enriched using collagen type IV. However, hDSPCs gradually lose their stem cell properties as in vitro passages continue. In the present study, we developed optimized in vitro culture condition to improve the stemness of these hDSPCs. To evaluate whether the stemness of hDSPCs is well sustained in various culture conditions, we measured the expression levels of SOX2, NANOG, and S100B, which are well-known representative dermal progenitor markers. We observed that hDSPCs grown in three-dimensional (3D) culture condition had higher expression levels of those markers compared with hDSPCs grown in two-dimensional (2D) culture condition. Under the 3D culture condition, we further demonstrated that a high glucose (4.5 g/L) concentration enhanced the expression levels of the dermal progenitor markers, whereas O 2 concentration did not affect. We also found that skin-derived precursor (SKP) culture medium was the most effective, among various culture media, in increasing the dermal progenitor marker expression. We finally demonstrated that this optimized culture condition enhanced the expression level of human telomerase reverse transcriptase (hTERT), the proliferation, and the multipotency of hDSPCs, an important characteristic of stem cells. Taken together, these results suggested that this novel in vitro culture condition improves the stemness of hDSPCs.

show abstract

“…1 In the literature, oxygen concentrations < 5% have been described as hypoxic conditions. [1][2][3] Hypoxia can create a potentially lethal environment and limit cellular respiration and growth or, alternatively, enhance the production of specific extracellular matrix components and increase angiogenesis. 4 A hypoxic environment plays a substantial and critical role in many essential physiological and developmental pathways.…”

Section: Introductionmentioning

confidence: 99%

Tracking Hypoxic Signaling in Encapsulated Stem Cells

Sahai

McFarland²,

Skiles³

et al. 2012

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

Oxygen is not only a nutrient but also an important signaling molecule whose concentration can influence the fate of stem cells. This study details the development of a marker of hypoxic signaling for use with encapsulated cells. Testing of the marker was performed with adipose-derived stem cells (ADSCs) in two-dimensional (2D) and 3D culture conditions in varied oxygen environments. The cells were genetically modified with our hypoxia marker, which produces a red fluorescent protein (DsRed-DR), under the control of a hypoxia-responsive element (HRE) trimer. For 3D culture, ADSCs were encapsulated in poly(ethylene glycol)-based hydrogels. The hypoxia marker (termed HRE DsRed-DR) is built on a recombinant adenovirus and ADSCs infected with the marker will display red fluorescence when hypoxic signaling is active. This marker was not designed to measure local oxygen concentration but rather to show how a cell perceives its local oxygen concentration. ADSCs cultured in both 2D and 3D were exposed to 20% or 1% oxygen environments for 96 h. In 2D at 20% O 2 , the marker signal was not observed during the study period. In 1% O 2 , the fluorescent signal was first observed at 24 h, with maximum prevalence observed at 96 h as 59% -3% cells expressed the marker. In 3D, the signal was observed in both 1% and 20% O 2 . The onset of signal in 1% O 2 was observed at 4 h, reaching maximum prevalence at 96 h with 76% -4% cells expressing the marker. Interestingly, hypoxic signal was also observed in 20% O 2 , with 13% -3% cells showing positive marker signal after 96 h. The transcription factor subunit hypoxia inducible factor-1a was tracked in these cells over the same time period by immunostaining and western blot analysis. Immunostaining results in 2D correlated well with our marker at 72 h and 96 h, but 3D results did not correlate well. The western blotting results in 2D and 3D correlated well with the fluorescent marker. The HRE DsRed-DR virus can be used to track the onset of this response for encapsulated, mesenchymal stem cells. Due to the importance of hypoxic signaling in determination of stem cell differentiation, this marker could be a useful tool for the tissue engineering community.

show abstract

Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

Cited by 184 publications

References 33 publications

Effect of low oxygen tension on the biological characteristics of human bone marrow mesenchymal stem cells

Effect of low oxygen tension on the biological characteristics of human bone marrow mesenchymal stem cells

Novel In Vitro Culture Condition Improves the Stemness of Human Dermal Stem/Progenitor Cells

Tracking Hypoxic Signaling in Encapsulated Stem Cells

Contact Info

Product

Resources

About