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Low oxygen tension positively influences cardiomyocyte progenitor cell function.

van Oorschot AA, Smits AM, Pardali E, Doevendans PA, Goumans MJ - J. Cell. Mol. Med. (2011)

Bottom Line: Previously we observed that cardiomyocyte progenitor cells (hCMPCs) isolated from the human heart differentiate spontaneously into cardiomyocytes and vascular cells when transplanted after myocardial infarction (MI) in the ischemic heart.After MI, deprivation of oxygen is the first major change in the cardiac environment.Knockdown of TSP-2 resulted in increased proliferation, migration and MMP activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Cell Biology and Center for Biomedical Genetics, Leiden University Medical Center, Leiden, The Netherlands.

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Long-term exposure to hypoxia inhibits CMPC migration via increased expression of the ECM modulator TSP-2. (A) The expression of MMP-2 in cells was determined via RT-qPCR and was not influenced by hypoxic conditions. (B) Secretion of MMP-2 in CM was demonstrated by zymography. The expression TIMP-1 (C), TIMP-2 (D), TSP-1 (E) and TSP-2 (F) was measured by RT-qPCR. (G) Secretion of TSP-1 and TSP-2 is increased when hCMPCs are exposed to hypoxia for 9 days. (H) After 9 days of culture, migration in a scratch assay was determined, and showed no difference between hypoxia and normoxia. For all genes, expression was normalized to day 0 and β-actin was measured as a housekeeping gene. #Significant difference compared to all groups; *significant difference between two groups.
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fig06: Long-term exposure to hypoxia inhibits CMPC migration via increased expression of the ECM modulator TSP-2. (A) The expression of MMP-2 in cells was determined via RT-qPCR and was not influenced by hypoxic conditions. (B) Secretion of MMP-2 in CM was demonstrated by zymography. The expression TIMP-1 (C), TIMP-2 (D), TSP-1 (E) and TSP-2 (F) was measured by RT-qPCR. (G) Secretion of TSP-1 and TSP-2 is increased when hCMPCs are exposed to hypoxia for 9 days. (H) After 9 days of culture, migration in a scratch assay was determined, and showed no difference between hypoxia and normoxia. For all genes, expression was normalized to day 0 and β-actin was measured as a housekeeping gene. #Significant difference compared to all groups; *significant difference between two groups.

Mentions: Because repair of ischemic damage of cardiac tissue takes longer than 1 day we also explored the effects of prolonged (6 and 9 days) exposure to hypoxia on the different aspects of migration. Because ECM modulators are important for the migration of (progenitor) cells through cardiac tissue, we determined the effect hypoxia had of MMP-2 and -9, and the MMP inhibitors TIMP and TSP [27, 28]. Using zymography and RT-qPCR we observed that hCMPCs express high levels MMP-2 mRNA and secreted large amounts of MMP-2 protein (Fig. 6A and B). Culturing hCMPCs under hypoxic conditions did not affect the MMP-2 gene expression nor the MMP-2 protein secretion (Fig. 6A and B). MMP-9 protein or mRNA levels were below the detection level in all conditions analysed (data not shown).


Low oxygen tension positively influences cardiomyocyte progenitor cell function.

van Oorschot AA, Smits AM, Pardali E, Doevendans PA, Goumans MJ - J. Cell. Mol. Med. (2011)

Long-term exposure to hypoxia inhibits CMPC migration via increased expression of the ECM modulator TSP-2. (A) The expression of MMP-2 in cells was determined via RT-qPCR and was not influenced by hypoxic conditions. (B) Secretion of MMP-2 in CM was demonstrated by zymography. The expression TIMP-1 (C), TIMP-2 (D), TSP-1 (E) and TSP-2 (F) was measured by RT-qPCR. (G) Secretion of TSP-1 and TSP-2 is increased when hCMPCs are exposed to hypoxia for 9 days. (H) After 9 days of culture, migration in a scratch assay was determined, and showed no difference between hypoxia and normoxia. For all genes, expression was normalized to day 0 and β-actin was measured as a housekeeping gene. #Significant difference compared to all groups; *significant difference between two groups.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4373441&req=5

fig06: Long-term exposure to hypoxia inhibits CMPC migration via increased expression of the ECM modulator TSP-2. (A) The expression of MMP-2 in cells was determined via RT-qPCR and was not influenced by hypoxic conditions. (B) Secretion of MMP-2 in CM was demonstrated by zymography. The expression TIMP-1 (C), TIMP-2 (D), TSP-1 (E) and TSP-2 (F) was measured by RT-qPCR. (G) Secretion of TSP-1 and TSP-2 is increased when hCMPCs are exposed to hypoxia for 9 days. (H) After 9 days of culture, migration in a scratch assay was determined, and showed no difference between hypoxia and normoxia. For all genes, expression was normalized to day 0 and β-actin was measured as a housekeeping gene. #Significant difference compared to all groups; *significant difference between two groups.
Mentions: Because repair of ischemic damage of cardiac tissue takes longer than 1 day we also explored the effects of prolonged (6 and 9 days) exposure to hypoxia on the different aspects of migration. Because ECM modulators are important for the migration of (progenitor) cells through cardiac tissue, we determined the effect hypoxia had of MMP-2 and -9, and the MMP inhibitors TIMP and TSP [27, 28]. Using zymography and RT-qPCR we observed that hCMPCs express high levels MMP-2 mRNA and secreted large amounts of MMP-2 protein (Fig. 6A and B). Culturing hCMPCs under hypoxic conditions did not affect the MMP-2 gene expression nor the MMP-2 protein secretion (Fig. 6A and B). MMP-9 protein or mRNA levels were below the detection level in all conditions analysed (data not shown).

Bottom Line: Previously we observed that cardiomyocyte progenitor cells (hCMPCs) isolated from the human heart differentiate spontaneously into cardiomyocytes and vascular cells when transplanted after myocardial infarction (MI) in the ischemic heart.After MI, deprivation of oxygen is the first major change in the cardiac environment.Knockdown of TSP-2 resulted in increased proliferation, migration and MMP activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Cell Biology and Center for Biomedical Genetics, Leiden University Medical Center, Leiden, The Netherlands.

Show MeSH
Related in: MedlinePlus