세포 외 pH 변화가 인간 골수 기질세포의 증식 및 분화에 미치는 영향
The Effects of Extracellular pH on Proliferation and Differentiation of human Bone Marrow Stem Cells
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연구목적: 본 연구는 세포 외 pH 변화에 따른 골수 기질세포의 증식 및 분화에 어떠한 영향을 미치는 지와 그와 관련된기전을 알아보았다. 연구방법: 본 연구의 가설을 검증하기 위해 HCl과 NaOH를 이용하여 배양액의 pH를 6.4에서 8.0의 다양한 범위를 유도하여 골수 기질세포의 증식 및 분화를 확인하였다. 또한, 관련 기전을 알아보기 위해 CaSR 및 ERK의 활성 억제를 통해골수 기질세포의 증식을 관찰하였다. 또한, 분화능과 관련하여 골 형성 관련 유전자인 Runx2, ALP, 및 OCN의 mRNA 발현 양상과 ALP 활성을 관찰하였다. 결 과: 골수 기질세포의 증식은 pH 7.6~8.0에서 통계적으로 유의하게 증가함을 보였으며, 이러한 변화는 CaSR/ERK pathway와 관련이 있음을 보였다. 반면에, pH 7.6 혹은 8.0에서는 감소된 ALP 및 OCN mRNA 발현 감소와 ALP 활성의감소가 관찰되었다. 결 론: 골수 기질세포의 증식 및 분화 능력은 다양한 요인에 의해 변화될 수 있는데, 그 중 세포 외 pH 변화에 따른알칼리 및 산성 환경도 하나의 중요한 요인으로 작용될 수 있음을 시사한다.
Abstract
Objectives: The purpose of this study is to identify whether the change of pH affects the proliferation and the differentiation of human bone marrow stem cells (hBMSCs) and what mechanism is underlied. Methods: To achieve objective of this study, hBMSCs were cultivated in the conditioned media adjusted to potential of hydrogen (pH) ranging from 6.4 to 8.0 using addition of hydrochloric acid (HCl) and sodium hydroxide (NaOH). The ratio of proliferation of hBMSCs according to the change of pH was measured for 24 h, 48 h, and 72 h using water-soluble tetrazolium salt (WST)-8 method. To elucidate the mechanism involved, hBMSCs was subjected to blocking extracellular signal-regulated kinases (ERK) and calcium sensing receptor (CaSR) activation. The Osteogenicrelated genes and alkaline phosphatase (ALP) activity were tested under the conditioned media. Results: The proliferation of hBMSCs was promoted under extracellular alkali conditions (pH 7.6~8.0) via CaSR/ERK pathway. On the other hand, the differentiation was inhibited/delayed via decreased ALP activity besides gene expression at pH 8.0. Conclusion: Extracellular alkali or acidic surrounding according to pH alteration can play a crucial role in hBMSC behavior including the proliferation and the differentiation.
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