Growth and stretch response of human exstrophy bladder smooth muscle cells: Molecular evidence of normal intrinsic function

Anna Orsola, Carlos R. Estrada, Hiep T. Nguyen, Alan B. Retik, Michael R. Freeman, Craig A Peters, Rosalyn M. Adam

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

OBJECTIVE: To establish primary cultures of smooth muscle cells (SMC) from human exstrophic bladders (E-SMC), and determine their in vitro growth dynamics and responses to mechanical stretch. MATERIALS AND METHODS: Primary cultures of E-SMC from three patients were established from exstrophic bladder tissue using an explant method. Growth dynamics were assessed using tetrazolium-dye uptake. The DNA synthesis rate in response to cyclic stretch-relaxation was determined with thymidine-incorporation assays. Expression of the SMC mitogen heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA in response to mechanical stretch was determined using semiquantitative reverse transcription-polymerase chain reaction. RESULTS: The approximate doubling time of the E-SMC grown in the presence of serum was 4 days, consistent with growth rates of SMC reported previously. E-SMC exposed to stretch had greater DNA synthesis, albeit to a lesser extent than previously seen with non-exstrophic SMC. The expression of HB-EGF was also increased in cells exposed to mechanical stimuli, consistent with our previous finding of stretch-regulated HB-EGF gene expression in bladder SMC. CONCLUSIONS: E-SMC had growth characteristics similar to those previously reported in non-exstrophic cells. E-SMC also had stretch-induced expression of HB-EGF mRNA. These observations provide evidence that despite development in an abnormal defunctionalized state, E-SMC retain the potential for normal growth, and may modulate this response through mechanisms similar to those operating in normal bladder SMC.

Original languageEnglish (US)
Pages (from-to)144-148
Number of pages5
JournalBJU International
Volume95
Issue number1
DOIs
StatePublished - Jan 1 2005

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Bladder Exstrophy
Smooth Muscle Myocytes
Growth
Epidermal Growth Factor
Heparin
Intercellular Signaling Peptides and Proteins
Urinary Bladder
Messenger RNA
DNA
Mitogens
Thymidine

Keywords

  • Bladder exstrophy
  • HB-EGF
  • Primary culture
  • Smooth muscle
  • Stretch

ASJC Scopus subject areas

  • Urology

Cite this

Growth and stretch response of human exstrophy bladder smooth muscle cells : Molecular evidence of normal intrinsic function. / Orsola, Anna; Estrada, Carlos R.; Nguyen, Hiep T.; Retik, Alan B.; Freeman, Michael R.; Peters, Craig A; Adam, Rosalyn M.

In: BJU International, Vol. 95, No. 1, 01.01.2005, p. 144-148.

Research output: Contribution to journalArticle

Orsola, Anna ; Estrada, Carlos R. ; Nguyen, Hiep T. ; Retik, Alan B. ; Freeman, Michael R. ; Peters, Craig A ; Adam, Rosalyn M. / Growth and stretch response of human exstrophy bladder smooth muscle cells : Molecular evidence of normal intrinsic function. In: BJU International. 2005 ; Vol. 95, No. 1. pp. 144-148.
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abstract = "OBJECTIVE: To establish primary cultures of smooth muscle cells (SMC) from human exstrophic bladders (E-SMC), and determine their in vitro growth dynamics and responses to mechanical stretch. MATERIALS AND METHODS: Primary cultures of E-SMC from three patients were established from exstrophic bladder tissue using an explant method. Growth dynamics were assessed using tetrazolium-dye uptake. The DNA synthesis rate in response to cyclic stretch-relaxation was determined with thymidine-incorporation assays. Expression of the SMC mitogen heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA in response to mechanical stretch was determined using semiquantitative reverse transcription-polymerase chain reaction. RESULTS: The approximate doubling time of the E-SMC grown in the presence of serum was 4 days, consistent with growth rates of SMC reported previously. E-SMC exposed to stretch had greater DNA synthesis, albeit to a lesser extent than previously seen with non-exstrophic SMC. The expression of HB-EGF was also increased in cells exposed to mechanical stimuli, consistent with our previous finding of stretch-regulated HB-EGF gene expression in bladder SMC. CONCLUSIONS: E-SMC had growth characteristics similar to those previously reported in non-exstrophic cells. E-SMC also had stretch-induced expression of HB-EGF mRNA. These observations provide evidence that despite development in an abnormal defunctionalized state, E-SMC retain the potential for normal growth, and may modulate this response through mechanisms similar to those operating in normal bladder SMC.",
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AU - Estrada, Carlos R.

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AU - Retik, Alan B.

AU - Freeman, Michael R.

AU - Peters, Craig A

AU - Adam, Rosalyn M.

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N2 - OBJECTIVE: To establish primary cultures of smooth muscle cells (SMC) from human exstrophic bladders (E-SMC), and determine their in vitro growth dynamics and responses to mechanical stretch. MATERIALS AND METHODS: Primary cultures of E-SMC from three patients were established from exstrophic bladder tissue using an explant method. Growth dynamics were assessed using tetrazolium-dye uptake. The DNA synthesis rate in response to cyclic stretch-relaxation was determined with thymidine-incorporation assays. Expression of the SMC mitogen heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA in response to mechanical stretch was determined using semiquantitative reverse transcription-polymerase chain reaction. RESULTS: The approximate doubling time of the E-SMC grown in the presence of serum was 4 days, consistent with growth rates of SMC reported previously. E-SMC exposed to stretch had greater DNA synthesis, albeit to a lesser extent than previously seen with non-exstrophic SMC. The expression of HB-EGF was also increased in cells exposed to mechanical stimuli, consistent with our previous finding of stretch-regulated HB-EGF gene expression in bladder SMC. CONCLUSIONS: E-SMC had growth characteristics similar to those previously reported in non-exstrophic cells. E-SMC also had stretch-induced expression of HB-EGF mRNA. These observations provide evidence that despite development in an abnormal defunctionalized state, E-SMC retain the potential for normal growth, and may modulate this response through mechanisms similar to those operating in normal bladder SMC.

AB - OBJECTIVE: To establish primary cultures of smooth muscle cells (SMC) from human exstrophic bladders (E-SMC), and determine their in vitro growth dynamics and responses to mechanical stretch. MATERIALS AND METHODS: Primary cultures of E-SMC from three patients were established from exstrophic bladder tissue using an explant method. Growth dynamics were assessed using tetrazolium-dye uptake. The DNA synthesis rate in response to cyclic stretch-relaxation was determined with thymidine-incorporation assays. Expression of the SMC mitogen heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA in response to mechanical stretch was determined using semiquantitative reverse transcription-polymerase chain reaction. RESULTS: The approximate doubling time of the E-SMC grown in the presence of serum was 4 days, consistent with growth rates of SMC reported previously. E-SMC exposed to stretch had greater DNA synthesis, albeit to a lesser extent than previously seen with non-exstrophic SMC. The expression of HB-EGF was also increased in cells exposed to mechanical stimuli, consistent with our previous finding of stretch-regulated HB-EGF gene expression in bladder SMC. CONCLUSIONS: E-SMC had growth characteristics similar to those previously reported in non-exstrophic cells. E-SMC also had stretch-induced expression of HB-EGF mRNA. These observations provide evidence that despite development in an abnormal defunctionalized state, E-SMC retain the potential for normal growth, and may modulate this response through mechanisms similar to those operating in normal bladder SMC.

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