Enhanced Healing of Rat Calvarial Bone Defects with Hypoxic Conditioned Medium from Mesenchymal Stem Cells through Increased Endogenous Stem Cell Migration via Regulation of ICAM-1 Targeted-microRNA-221

Chang, Woochul,Kim, Ran,Park, Sang In,Jung, Yu Jin,Ham, Onju,Lee, Jihyun,Kim, Ji Hyeong,Oh, Sekyung,Lee, Min Young,Kim, Jongmin,Park, Moon-Seo,Chung, Yong-An,Hwang, Ki-Chul,Maeng, Lee-So Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.7

The use of conditioned medium from mesenchymal stem cells may be a feasible approach for regeneration of bone defects through secretion of various components of mesenchymal stem cells such as cytokines, chemokines, and growth factors. Mesenchymal stem cells secrete and accumulate multiple factors in conditioned medium under specific physiological conditions. In this study, we investigated whether the conditioned medium collected under hypoxic condition could effectively influence bone regeneration through enhanced migration and adhesion of endogenous mesenchymal stem cells. Cell migration and adhesion abilities were increased through overexpression of intercellular adhesion molecule-1 in hypoxic conditioned medium treated group. Intercellular adhesion molecule-1 was upregulated by microRNA-221 in mesenchymal stem cells because microRNAs are key regulators of various biological functions via gene expression. To investigate the effects in vivo, evaluation of bone regeneration by computed tomography and histological assays revealed that osteogenesis was enhanced in the hypoxic conditioned medium group relative to the other groups. These results suggest that behavioral changes of endogenous mesenchymal stem cells through microRNA-221 targeted-intercellular adhesion molecule-1 expression under hypoxic conditions may be a potential treatment for patients with bone defects.

Enhanced Healing of Rat Calvarial Bone Defects with Hypoxic Conditioned Medium from Mesenchymal Stem Cells through Increased Endogenous Stem Cell Migration via Regulation of ICAM-1 TargetedmicroRNA- 221

장우철,이소명,김란,박상인,정유진,함온주,이지현,김지형,오세경,이민영,김종민,박문서,정용안,황기철,맹이소 한국분자세포생물학회 2015 Molecules and cells Vol.38 No.7

The use of conditioned medium from mesenchymal stem cells may be a feasible approach for regeneration of bone defects through secretion of various components of mesenchymal stem cells such as cytokines, chemokines, and growth factors. Mesenchymal stem cells secrete and accumulate multiple factors in conditioned medium under specific physiological conditions. In this study, we investigated whether the conditioned medium collected under hypoxic condition could effectively influence bone regeneration through enhanced migration and adhesion of endogenous mesenchymal stem cells. Cell migration and adhesion abilities were increased through overexpression of intercellular adhesion molecule-1 in hypoxic conditioned medium treated group. Intercellular adhesion molecule- 1 was upregulated by microRNA-221 in mesenchymalstem cells because microRNAs are key regulators of various biological functions via gene expression. To investigate the effects in vivo, evaluation of bone regeneration by computed tomography and histological assays revealed that osteogenesis was enhanced in the hypoxic conditioned medium group relative to the other groups. These results suggest that behavioral changes of endogenous mesenchymal stem cells through microRNA-221 targetedintercellular adhesion molecule-1 expression under hypoxic conditions may be a potential treatment for patients with bone defects.

『高峰和尙禪要』에 나타난 華嚴的 考察

김숙현(修鏡) 한국선학회 2011 한국선학 Vol.30 No.-

『선요』는 간화선수행의 지침서로서 교학사상을 살펴볼 수 있는 대목은 그리 많지 않다. 하지만 화엄교학과 간화선이 이론과 실천의 관계를 가진다고 할 정도로 화엄사상이 간화선에 미친 영향은 크다고 알려져 왔다. 『선요』를 화엄적 안목에서 고찰해보고자 시도한 내용은「開堂普說」(제1)과 「解題示衆」(제19)이다. 이는 화엄사상이 저변에 자리하고 있다고 보았기 때문이다. 고찰의 방법은「開堂普說」의 경우 방거사의 게송(全4句)에 관한 고봉의 풀이를 性起論(제1,2구)과 緣起論(제3,4구)으로 나누고, 禪問答은 舊來成佛論으로 구분하여 고찰하였다.「解題示衆」은 인용문에 대한 출처를 밝히고, 또 화엄사상과 관련된 사상이 무엇인지 간략하게 살펴보았다. Ⅱ.「개당보설」에 나타난 화엄사상에서, 첫째 性起論에 해당되는 제1구는 방거사가 우주 현상계의 모습을 있는 그대로 표현한 대목이라면 고봉은 뱀과 용, 범부와 성인이라는 비유를 통해 외형적으로는 다른 개체들이 함께 어울려 사는 세상이지만, 근원적으로는 同質本性의 존재이기 때문에 다 함께 모여 살 수 있다는 法界觀을 드러내었다. 제2구에서 고봉은 은유법을 사용하여 무위법을 배우는 중생들이 佛祖와 대등한 자이며, 세상을 꿰뚫는 지혜를 가지고 있음을 밝혔다. 이것은 앞의 제1구가 총체적인 측면에서 法界相을 보여준 것이라면 제2구는 法界性을 말하고 있는 것으로 性起論에 해당된다. 다시 말하면 이 세상이 성기에 의존하고 있는 법계임을 밝힌 내용이다. 그러므로 여기서는 모든 존재가 성기의 顯現으로서 누구나 닦기만 하면 성불할 수 있다는 가능성을 제시하였다고 하겠다. 둘째 緣起論에 해당되는 제3구에서 ‘동서10만 남북8천’이란 세상 어느 곳이든 選佛場이라는 공간적 개념과 縱橫無盡하고 殺活自在한 경지라는 사상적 측면도 포함하고 있다. 선불장 아닌 곳이 없는 이 세계가 바로 화엄에서 말하는 事事無碍法界임을 나타내고 있는 말이다. 제4구는 성불할 수 있는 근거로서 연기를 밝히고 있는 내용이다. ‘옛길이 드러난다(揚古路)’는 것은 본분 또는 성기가 드러남을 뜻한다. 성기가 드러나는 것은 마음을 비우는 淨緣起에 의해 가능하다. 성기는 연기에 의해 체득되어지기 때문이다. 그러므로 고봉은 일체상이 연기로부터 일어난 줄 알며, 연기는 제 성품이 없어서(緣起無性) 성기를 의지하고 있는 줄을 체득하여 일체상이 곧 성기의 현현임을 깨닫는다면 절대로 二乘처럼 공에 떨어지지 않는다하였다. 마지막 구절은 성기로부터 生起한 존재가 정연기 수행을 통해 다시 성기로 돌아가는 것을 묘사하였다. 셋째 舊來成佛論은 고봉과 승려 간에 오고 간 선문답의 내용이다. 고봉은 초목과 無情에 이르기까지 구래성불이라는 입장을 밝히고 있으며, 이러한 구래성불은 성기사상이 토대가 된다. Ⅲ.「解題示衆」에 인용된 華嚴經句에서는「如來出現品」과「夜摩天宮菩薩說偈品」의 내용을 직접 인용하여 이를 주제로 설법하고 있다.「여래출현품」의 인용문에서는 성기사상을 엿볼 수 있으며, 「야마천궁보살설게품」의 인용문에서는 ‘一心’에 관한 것을 말하고자 한 것이라 생각된다. 이상의 고찰을 통해 고봉은 선사의 입장에서 늘 ‘무엇을 깨닫고 어떻게 수행할 것인가’에 초점이 맞추어져 있기 하지만 선수행의 이론적 측면에서는 화엄교학이 자리하고 있음을 알 수 있다. Chanyao as a guide book of kanhua Chan(Kor. Ganhwaseon) practice does not contain many related doctrines. It is, however, well known that the huayan(Kor. hwaeom) philosophy had an influence on Ganhwaseon so greatly that the former provides a theoretical basis of the latter. The author examines the perspective of hwaeom philosophy, which underlies Chanyao, specifically in the 1st section General Opening Sermon「開堂普說」 and the 19th Instruction on the Title of the Sutra「解題示衆」. The method of the research is done as follows: in the case of General Opening Sermon, the author divided Gaofeng Yuanmiao’s 高峰原妙 explanation of Householder Pang’s verse into two, ie., the theories of arising from the original nature性起論(the first and second lines) and from conditions緣起論(the third and fourth lines), and then Seon encounter dialogues into the theory of attaining buddhahood every moment舊來成佛論; in case of Instruction on the Title of the Sutra, the author made it clear where the excerpts come from and also briefly studied in what form the hwaeom thought is contained. To be more specific, the first line in Pang’s verse describes the phenomenal world as it is whereas Gaofeng, through an analogy with the serpent and dragon, and the ordinary man and saint, reveals the perspective of the dharma-realm法界觀 in which diverse creatures, sharing the same fundamental inherent nature with each other, can live in harmony altogether despite their seeming difference in appearance. In the second phrase, Gaofeng uses a metaphor and brings it to light that the sentient beings learning unconditioned existence無爲法 are equal to the buddhas and patriarchs, and have a penetrating wisdom. This reflects the theory of arising from the original nature while the first line manifests phenomena of the dharma realm法界相 on the whole. In other words, it unfolds that the world is a dharma realm dependent on the arising of the original nature. Thus, it can be said that anyone has the potential to attain buddhahood if only to be engaged in cultivation since each of them is a manifestation of the arising of the original nature. In the third line, Gaofeng’s ‘The east-west distance is one hundred thousand ri and the north-south, eighty thousand ri,’ contains the meaning that spatially there is nowhere but can be a place for selecting the buddha and ideologically it expresses the spiritual stage of neither hindrance nor life and death. That is to say, the world as a place for selecting the buddha anywhere is the realm of non-obstruction between phenomena事事無碍法界. The fourth phrase reveals conditioned arising緣起 as a basis for achieving buddhahood. The line of ‘An old path reveals itself’ means that buddha-nature or original reality manifests itself. This stage is possible when one reaches conditioned arising of purity淨緣起. After all, the stage of arising from the original nature성기 can be attained through that of conditioned arising. Therefore, Gaofeng states that, if one understands all phenomena resulting from conditioned arising, and this conditioned arising relies upon arising from the original nature, one will never fall into voidness空 as the two vehicles二乘 did. Gaofeng’s last line describes that the being who has arisen from the original nature return to the original nature through the practice of conditioned arising of purity. Third, the theory of attaining buddhahood every moment舊來成佛論 is contained in the Chan encounter dialogue between Gaofeng and other monks. Master Gaofeng takes the stance that all things including insentient things like plants and trees attained buddhahood already and this theory lays a foundation of the thought of arising from the original nature. In the part of Instruction on the Title of the Sutra, Gaofeng directly cites some excerpts from Chapter on the Appearance of the Tathāgata「如來出現品」 and Ascending to the Suyama Heaven Palace「夜摩天宮菩薩說偈品」 of the Huayan jing. The author think that the citations are used for the purpose of introducing the thought of arising from the original nature and one mind一心. From the investigation so far, the author understand that Gaofeng, as a Chan master, always focuses on ‘What to realize and how to practice?’ but theoretically he applies the huayan philosophy to his practice.

고타마 싯닷타는 어떻게 붓다가 되었나?

박태원(Park, Tae-Won) 새한철학회 2017 哲學論叢 Vol.88 No.2

붓다가 일러주는 것처럼, 모든 현상은 그것을 발생시키는 조건들에 의해 발생한다. 현상을 볼 때는, 그것을 발생시킨 원인조건들을 동시에 고려해야 진실에 접근할 수 있고, 또 문제를 제대로 풀 수 있다. 실존의 근원적이고도 궁극적인 문제를 풀려는 청년 고타마 싯닷타의 모험적 도전이 성공할 수 있었던 것도, 성공할 수 있는 조건들을 확보하였기 때문이다. 고타마 싯닷타의 수행과정을 주목한다는 것은, 깨달음의 발생과 인과적 연관을 맺는 조건들이 무엇인가를 읽어내려는 것이다. 붓다가 사람들에게 자신의 수행과정을 회고해준 것도, 자신이 성공할 수 있었던 요인들, 그 ‘깨달음의 발생조건들’을, 체험적 증언을 통해 알려주려는 것이었을 것이다. 제자들의 기억과 후인들의 전승을 통해 전해지고 있는 붓다의 수행 회고담에서 고타마 싯닷타의 성공 요인 네 가지를 골라 그 철학적 의미를 읽어본다. 고타마 싯닷타의 수행과정은 니까야/아함 문헌에서 두 유형으로 전해진다. 붓다가 자신의 수행경험을 회고하는 유형이 하나이고, 자신의 수행과정을 구도자의 표준 모델로 일반화시켜 설하는 유형이 다른 하나이다. 특히 구조와 내용에서 정합성을 보여주는 일반화 유형이 자주 설해지고 있다는 것은, 붓다가 자신의 깨달음을 세상과 공유하기 위해 수행과정의 경험을 적극적으로 활용했음을 의미한다. 니까야/아함이 전하는 내용을 종합해 볼 때, 4가지 조건들을 주목해야 할 것으로 생각한다. 〈1) ‘이해/언어’와 ‘탈(脫)이해/탈(脫)언어’의 차이 및 관계에 대한 개안, 2) 조건적 발생에 대한 개안 – 이지적 연기 깨달음, 3) 모든 조건과 경험에 갇히지 않는 능력의 확보 – 새로운 선법(禪法)의 수립, 4) ‘조건인과적 발생’에 대한 직접지(直接知) 성취 – 체득적 연기 깨달음〉이 그것이다. The practice-process of Gotama Siddhattha is described as two kinds of form in Nikāya/Agama. One is the form of Buddha’s recollections of his own, the other is the form of general mode on practice of all practicers. And the form of general mode which show a coherence in structure and contents appears more frequently. This means that Buddha used his experience of practice actively to enlighten his disciples in common. Four factors of success are to be appointed. They are 1) Gaining eyesight on the diffe rence and relation between ‘intellectual/verbal understanding’ and ‘non-intellectual/verbal understanding’, 2) Gaining eyesight on the Conditioned-occurrence –an intellectual enlightenment on Conditioned-occurrence(緣起), 3) Securing the capacity not being confined in any conditions and experiences –establishment of a new Jhana-practice(禪法), 4) Accomplishing the direct-understanding(直接知) on Conditioned-occurrence(緣起) - experiential enlightenment on Conditioned-occurrence(緣起). Gotama Siddhattha‘s accomplishment of the insight on the Conditioned-occurrence and the capacity not being confined in any conditions and experiences is a condition that can accomplish a new evolution of Human. Gotama Siddhattha‘s success gives a prospect upon the human who does not make any criteria unconditioned and absolute, who is flexible on the modification, supplementation, abandonment of one’s criteria, who can make relationship with differences peacefully and mutual-beneficially.

Identification of proteins associated with cell repair from canine adipose derived mesenchymal stem cells isolated conditioned media

Ahmad Yar Qamar,Xun Fang,Jongki Cho 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

The objective of this study was to identify the proteins actively involved in the protection and repair of damaged cells, secreted by canine adipose derived mesenchymal stem cells (AT-MSCs) into the conditioned media. For this purpose, conditioned media (CM) was recovered from passage three stage canine AT-MSCs and skin fibroblasts cultured in serum free media after 24, 48 and 72 h. The extraction of exosomes was performed from 10-20 ml of CM using total exosome isolation kit. The isolated exosomes were then subjected to western analysis for the identification of annexin-I, annexin-II, histone H3 and dysferlin proteins. Results demonstrated the expression of proteins in the conditioned media isolated from canine AT-MSCs reflecting their potential in reducing the extent of damage at cellular levels. In conclusion, the conditioned media derived from canine AT-MSCs can be helpful in restoring the normal structure of cells both in vivo and in vitro conditions.

Studies on Conditioned Media in Human Cells: Evaluation Using Various Cell and Culture Conditions, Animal Disease Models

김근천,이은주 사단법인 한국동물생명공학회 2018 한국동물생명공학회지 Vol.33 No.1

In the last several decades, cell therapy research has increased worldwide. Many studies have been conducted on cell therapy, and have revealed that transplanted cells did not survive for long, and implanted cells remained inactive causing immune rejection depending on the patient’s condition. Therefore, studies on cell-free therapy need to be conducted. To overcome these limitations, an alternative is the use of supernatant from cells, called “conditioned media (CM).” During in vitro cell culture, culture media supply nutrients to maintain cell characteristics and viability. In the culture, cells not only consume nutrients but also release beneficial proteins and substances, which are called “secretome.” CM from cells can be stored for a long time and is easy to handle. Moreover, secretome in CM can also be measured; exact amount of secretome is important to set the standard value for disease treatment. Here, we reviewed studies on CM and confirmed that various secretomes from CM were identified in these studies. Moreover, these findings could benefit cell and animal studies in future. In conclusion, CM could be a potential candidate for an alternative to cell therapy.

Studies on Conditioned Media in Human Cells: Evaluation Using Various Cell and Culture Conditions, Animal Disease Models

Kim, Keun Cheon,Lee, Eun Ju The Korean Society of Embryo Transfer 2018 한국동물생명공학회지 Vol.33 No.1

In the last several decades, cell therapy research has increased worldwide. Many studies have been conducted on cell therapy, and have revealed that transplanted cells did not survive for long, and implanted cells remained inactive causing immune rejection depending on the patient's condition. Therefore, studies on cell-free therapy need to be conducted. To overcome these limitations, an alternative is the use of supernatant from cells, called "conditioned media (CM)." During in vitro cell culture, culture media supply nutrients to maintain cell characteristics and viability. In the culture, cells not only consume nutrients but also release beneficial proteins and substances, which are called "secretome." CM from cells can be stored for a long time and is easy to handle. Moreover, secretome in CM can also be measured; exact amount of secretome is important to set the standard value for disease treatment. Here, we reviewed studies on CM and confirmed that various secretomes from CM were identified in these studies. Moreover, these findings could benefit cell and animal studies in future. In conclusion, CM could be a potential candidate for an alternative to cell therapy.

Studies on Conditioned Media in Human Cells: Evaluation Using Various Cell and Culture Conditions, Animal Disease Models

Keun Cheon Kim,Eun Ju Lee 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.1

In the last several decades, cell therapy research has increased worldwide. Many studies have been conducted on cell therapy, and have revealed that transplanted cells did not survive for long, and implanted cells remained inactive causing immune rejection depending on the patient’s condition. Therefore, studies on cell-free therapy need to be conducted. To overcome these limitations, an alternative is the use of supernatant from cells, called “conditioned media (CM).” During in vitro cell culture, culture media supply nutrients to maintain cell characteristics and viability. In the culture, cells not only consume nutrients but also release beneficial proteins and substances, which are called “secretome.” CM from cells can be stored for a long time and is easy to handle. Moreover, secretome in CM can also be measured; exact amount of secretome is important to set the standard value for disease treatment. Here, we reviewed studies on CM and confirmed that various secretomes from CM were identified in these studies. Moreover, these findings could benefit cell and animal studies in future. In conclusion, CM could be a potential candidate for an alternative to cell therapy.

고온기 재배시설에 따른 털목이 ‘건이’의 생육 특성

김길자,김다미,안호섭,최진경,권오도 한국버섯학회 2019 한국버섯학회지 Vol.17 No.1

We assessed the growth characteristics of Auricularia polytricha ‘Geoni’ cultivated in a simple greenhouse constructed of polyethylene (PE) without air conditioning (high temperature) and in an air conditioned mushroomhouse. The successful cultivation of A. polytricha ‘Geoni’ at high temperatures can reduce energy and facility investment costs. The comparison of growth characteristics of the fungi grown under the different temperature conditions revealed that fruit bodies were larger in the higher temperature condition, but were brighter in the lower temperature condition. Additionally, fruit body physiology was found to be not affected by temperature. In the PE greenhouse, the fresh weight of fruit body was higher in mid-June and early July. Therefore, it was possible to effectively control the growth period of the mushrooms during a high-temperature period. The findings indicate the potential to cultivate A. polytricha ‘Geoni’in a simple PE greenhouse that is not cooled in summer, thus reducing energy costs.

Therapeutic potential of BMSC-conditioned medium in an in vitro model of renal fibrosis using the RPTEC/TERT1 cell line

( Yunji Kim ),( Dayeon Kang ),( Ga-eun Choi ),( Sang Dae Kim ),( Sun-ja Yang ),( Hyosang Kim ),( Dalsan You ),( Choung Soo Kim ),( Nayoung Suh ) 생화학분자생물학회 2024 BMB Reports Vol.57 No.2

We investigated the therapeutic potential of bone marrow-derived mesenchymal stem cell-conditioned medium (BMSC-CM) on immortalized renal proximal tubule epithelial cells (RPTEC/ TERT1) in a fibrotic environment. To replicate the increased stiffness characteristic of kidneys in chronic kidney disease, we utilized polyacrylamide gel platforms. A stiff matrix was shown to increase α-smooth muscle actin (α-SMA) levels, indicating fibrogenic activation in RPTEC/TERT1 cells. Interestingly, treatment with BMSC-CM resulted in significant reductions in the levels of fibrotic markers (α-SMA and vimentin) and increases in the levels of the epithelial marker E-cadherin and aquaporin 7, particularly under stiff conditions. Furthermore, BMSC-CM modified microRNA (miRNA) expression and reduced oxidative stress levels in these cells. Our findings suggest that BMSC-CM can modulate cellular morphology, miRNA expression, and oxidative stress in RPTEC/TERT1 cells, highlighting its therapeutic potential in fibrotic kidney disease.