Growth factors improve the proliferation of Jeju black pig muscle cells by regulating myogenic differentiation 1 and growth-related genes

Park, Jinryong,Lee, Jeongeun,Song, Ki-Duk,Kim, Sung-Jo,Kim, Dae Cheol,Lee, Sang Cheol,Son, Young June,Choi, Hyun Woo,Shim, Kwanseob 아세아태평양축산학회 2021 Animal Bioscience Vol.34 No.8

Objective: The growth rate of pigs is related to differentiation and proliferation of muscle cells, which are regulated by growth factors and expression of growth-related genes. Thus, the objective of this study was to establish optimal culture conditions for Jeju black pig (JBP) muscle cells and determine the relationship of various factors involved in muscle growth with the proliferation of JBP muscle cells. Methods: Muscles were taken from the femur skeletal muscle of JBP embryos. After isolation of the muscle cells, cells were cultured in a 6-well plate under four different culture conditions to optimize culture conditions for JBP muscle cells. To analyze proliferation rate of JBP muscle cells, these muscle cells were seeded into 6-well plates at a density of 1.5×10⁵ cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (MyoD) expression and growth-related gene expression in JBP muscle cells, respectively. Results: We established a muscle cell line from JBP embryos and optimized its culture conditions. These muscle cells were positive for MyoD, but not for paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (EGF+bFGF) than that without a growth factor or containing EGF alone. Treatment with EGF and bFGF significantly induced the expression of MyoD protein, an important transcription factor in muscle cells. Moreover, we checked the changes of expression of growth-related genes in JBP muscle cells by presence or absence of growth factors. Expression level of collagen type XXI alpha 1 gene was changed only when EGF and bFGF were added together to culture media for JBP muscle cells. Conclusion: Concurrent use of EGF and bFGF increased the expression of MyoD protein, thus regulating the proliferation of JBP muscle cells and the expression of growth-related genes.

배양된 쥐 갑상선세포 성장에 미치는 각종 성장인자들과 TSH 수용체항체간의 상호작용 및 그 기전에 관한 연구

조보연(Bo Youn Cho),송영기(Young Kee Shong),이홍규(Hong Kyu Lee),고창순(Chang Soon Koh),민헌기(Hun Ki Min) 대한내과학회 1989 대한내과학회지 Vol.37 No.3

N/A To evaluate the interaction of various growth factors and TSH receptor antibodies (TRAb) on the growth of cultured rat thyroid cells, FRTL-5 cells, and to verify their mechanisms of action, we measured 3H-thymidine incorporation into FRTL-5 cells with various combinations of TSH, Graves' IgG and other growth stimulators (insulin, IGF-I, multiplication stimulation activity, forskolin, dBcAMP and phorbol ester). We evaluated the effects of blocking TRAb and adenosine on the growth factor-stimulated growth of FRTL-5 cells. Insulin, IGF-I and MSA increased 3H-thymidine incorporation into FRTL-5 cells and showed a synergistic effect when incubated simultaneously with TSH or Graves IgG. Forskolin, PGE2, and dBcAMP also increased 3H-thymidine incorporation by themselves and their effects were synergistically potentiated by insulin and IGF-I. Adenosine inhibited both TSH and Graves' IgG induced 3H-thymidine incorporation into FRTL-5 cells. Blocking TRAb inhibited both TSH and Graves' IgG induced 3H-thymidine incorporation, but did not inhibit the effects of insulin, IGF-I, forskolin and dBcAMP on the growth of FRTL-5 cells. Phorbol ester (TPA) did stimulate the growth of FRTL-5 cells by itself and inhibited both TSH and Graves IgG induced 3H-thymidine incorporation into FRTL-5 cells. These results suggest that 1) there might be two or more signal transduction systems for the growth of thyroid cells; TSH and Graves IgG stimulate the growth of thyroid cells through the adenylate cyclase-cAMP system, and IGF-I and other growth facors act through another system, and 2) blocking TRAb may inhibit the growth of thyroid cells by inhibiting TSH-stimulated cAMP generation.

Effects of heparin-binding epidermal growth factor-like growth factor on cell repopulation and signal transduction in periodontal ligament cells after scratch wounding <i>in vitro</i>

Lee, J. S.,Kim, J. M.,Hong, E. K.,Kim, S.-O.,Yoo, Y.-J.,Cha, J.-H. Blackwell Publishing Ltd 2009 Journal of periodontal research Vol.44 No.1

<P>Background and Objective: </P><P>A growing amount of attention has been placed on periodontal regeneration and wound healing for periodontal therapy. This study was conducted in an effort to determine the effects of heparin-binding epidermal growth factor-like growth factor on cell repopulation and signal transduction in periodontal ligament cells after scratch wounding <I>in vitro</I>.</P><P>Material and Methods: </P><P>Human periodontal ligament cells were acquired from explant tissue of human healthy periodontal ligament. After the wounding of periodontal ligament cells, the change in expression of heparin-binding epidermal growth factor-like growth factor and epidermal growth factor receptors 1–4 mRNA was assessed. The effects of heparin-binding epidermal growth factor-like growth factor on periodontal ligament cell proliferation and repopulation were assessed <I>in vitro</I> via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and by photographing the injuries, respectively. Extracellular signal-regulated kinase (Erk)1/2, p38 and Akt phosphorylation was characterized via western blotting.</P><P>Results: </P><P>Scratch wounding resulted in a significant up-regulation of heparin-binding epidermal growth factor-like growth factor mRNA expression, whereas wounding had no effect on the expression levels of epidermal growth factor receptors 1–4. Interestingly, no expression of epidermal growth factor receptors 2 and 4 was detectable prior to or after wounding. Heparin-binding epidermal growth factor-like growth factor treatment promoted the proliferation and repopulation of periodontal ligament cells. The scratch wounding also stimulated the phosphorylation of Erk1/2 and p38, but not of Akt, in periodontal ligament cells, and heparin-binding epidermal growth factor-like growth factor treatment applied after wounding amplified and extended the activations of Erk1/2 and p38, but not of Akt. Furthermore, Erk1/2 inhibition blocked the process of cell repopulation induced by heparin-binding epidermal growth factor-like growth factor, whereas the inhibition of p38 delayed the process.</P><P>Conclusion: </P><P>These results indicate that heparin-binding epidermal growth factor-like growth factor may constitute a critical factor in the wound healing of human periodontal ligament cells by a mechanism that requires the activation of Erk1/2 via specific interaction with epidermal growth factor receptor 1.</P>

COX 억제제에 의해 유도되는 구강편평세포암종 세포주의 성장 억제 효과

박광진,한세진,이재훈,Park, Gwang-Jin,Han, Se-Jin,Lee, Jae-Hoon 대한악안면성형재건외과학회 2008 Maxillofacial Plastic Reconstructive Surgery Vol.30 No.4

The objectives of this study was to explore the growth pattern of the oral squamous cell carcinoma when overexpressed COX was inhibited, explore the pathway that COX inhibitors suppressed the proliferation of cancer cells, and then hereafter investigate the potential of COX as chemopreventive target for oral squamous cell carcinoma. For confirming the COX-dependent effect and mechanisms on growth of the oral cancer cells, we treated the nonselective NSAID, Mefenamic acid and COX-2 selective inhibitor, Celecoxib in HN4 cell line. And then the cell line was evaluated with MTT assay and growth curve, the production of PGE2, total RNA extraction and RT-PCR analysis, and TEM The results were obtained as follows: 1. After administration of medication, in the result of MTT assay, Celecoxib inoculated group inhibit the cell growth rather than Mefenamic acid inoculated group. 2. The growth curve of cell line showed as time passes by there was a dramatic cell growth in the control group, and gradual growth inhibition was found in medication inoculated group and, in Celecoxib inoculated group there was more inhibition of cell growth. 3. After the administration of medication, Celecoxib tend to inhibit the synthesis of PGE2 more than Mefenamic acid. Mefenamic acid inhibit the synthesis of PGE2 more as the concentration gets high, but Celecoxib inhibited the synthesis of PGE2 even in low concentration. 4. After the administration of medication, the revelation of COX mRNA in cell line, there was a 50% decrease in COX-1, 60% decrease in COX-2 as in $50{\mu}M$ Mefenamic acid, and in Celecoxib $50{\mu}M$ there was not much difference in COX-1 and 90% decrease in COX-2 was found. 5. HN4 cell line showed broken nucleus and tangled cytoskeleton bundles in cytoplasm which meant apoptotic features after the treatment of Celecoxib in TEM view. Depending on the above results, we estimate that the inhibition of the expression of COX-2 cause the growth suppression of the oral squamous cell carcinoma, and it get achieved through pathway of reduced PGE2 production and increased apoptosis. In addition to, because COX-2 selective inhibitor specifically act to COX-2, it is considered that COX-2 selective inhibitor has the adequate potential as chemopreventive agent for oral squamous cell carcinoma.

사람 중간엽줄기세포 성장에 미치는 basic fibroblast growth factor의 영항

김성수,최정원,곽규범,이영돈,서해영 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.6

Human mesenchymal stem cells (hMSCs) are multipotent stem cells that can differentiate into several mesenchymal lineage cells. In this study, we established conditions that allowed a long term expansion of hMSCs. To search for the optimum culture condition, growth rates of hMSCs were measured in the presence of several growth factors. Hepatic growth factor (HGF) and leukemia inhibitory factor (LIF) did not facilitate proliferation of hMSCs. In contrast, basic fibroblast growth factor (bFGF) effectively promoted growth of the cells in vitro by 3 fold. The growth stimulatory effect of bFGF was dependent on the concentration. The adipogenic potential was dramatically decreased in hMSCs isolated from an aged donor whereas osteogenic potential was minimally decreased. Addition of bFGF resumed the adipogenic and osteogenic differentiation potential. Thus, the cells that expanded in the presence of bFGF retained the potential to differentiate into adipogenic, chondrogenic, or osteogenic lineage cells. MSCs could be expanded for at least 8 passages with bFGF and the resulting cells retained the normal karyotype. The cells were positive for CD9, CD13, CD15, CD90, CD137, and CD140b; but negative for CD14, CD34, and CD45. Importantly, the cells were found to express a neural stem cell marker, nestin, and a neuronal marker, β-tubulin III. The results suggest that bFGF promote proliferation while maintaining multi-lineage differentiation potency of hMSCs. Finally, we suggest that it is critical to identify novel markers other than nestin or β-tubulin III to monitor acquisition of neuronal phenotypes by hMSCs. 중간엽줄기세포는 중배엽유래의 세포로 분화할 수 있는 다분화능을 갖는 줄기세포로서 골수에서 쉽게 채취할 수 있다. 본 연구에서는 사람의 중간엽줄기세포를 시험관내에서 장기간 배양시 세포의 증식과 다분화능에 있어서 basic fibroblast growth factor (bFGF)의 영향을 조사하였다.세포를 배양할 수 있는 최적의 조건을 설정하기 위하여 여러 성장인자를 시험하였다. Hepatic growth factor (HGF)와 leukemia inhibitory factor (LIF)는 골수에서 분리한 중간엽줄기세포의 증식에 별다른 영향을 주지 않았으나, bFGF는 시험관내에서 세포의 성장을 3배정도 증가시켰으며, 그 효과에 있어서 bFGF의 농도가 10 ng/ml 이상인 경우에는 큰 차이가 없었다. 저연령층의 골수에서 추출한 중간엽줄기세포는 bFGF의 존재와 상관없이 지방세포, 뼈세포, 연골세포로 분화하는 잠재력을 유지하고 있었으며 CD9, CD13, CD15, CD90, CD137, CD140b에 양성이었고, CD14, CD34, CD45에는 음성이었으며, 정상적인 핵형을 유지하고 있었다. 반면에 고령의 공여자로부터 분리한 중간엽줄기세포는 bFGF가 없을 경우 지방세포로의 분화능이 현격히 낮았으나, bFGF가 있는 경우에는 지방세포 및 뼈세포로의 분화능이 유지되었다. 또한 bFGF가 첨가된 배양액에서 증식시킨 중간엽줄기세포는 미분화상태에서도 이미 신경줄기세포의 표지자인 nestin과 신경세포의 표지자인 β-tubulin III을 발현하고 있었다. 이러한 결과는 bFGF가 사람의 중간엽줄기세포의 증식과 다분화능을 유지시키는데 중요한 성장인자로 작용함을 의미하고 있다. 또한, 중간엽줄기세포를 신경계질환의 세포치료제로서 사용하기 위해서는 미분화상태와 신경세포로 분화된 후를 구별할 수 있는 새로운 표식인자가 필요함을 제시하고 있다.

Hair-growth-promoting effect of conditioned medium of high integrin α6 and low CD 71 (α6bri/CD71dim) positive keratinocyte cells

( Young Jae Kim ),( Chong Hyun Won ),( Yun Mi Jeong ),( Sangjin Kang ),( Tae Sung Koo ),( So Hyun Park ),( Ki Young Park ),( Young Kwan Sung ),( Jong Hyuk Sung ) 대한피부과학회 2015 대한피부과학회 학술발표대회집 Vol.67 No.2

Background: Keratinocyte stem/progenitor cells (KSCs) reside in the bulge region of the hair follicles and may be involved in hair growth and regeneration. Objectives: We studied the hair growth promotion effects of KSC-CM on human HF organ culture and on a C3H/HeN mouse model. We then investigated the proliferative effect of KSC-CM on both human hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells. Methods: Hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells were treated with conditioned medium (CM) of KSCs. Moreover, the effects of KSC-CM on hair growth were examined ex vivo and in vivo. A human growth factor chip array and RT-PCR were employed to identify enriched proteins in KSC-CMas compared with CM from keratinocytes. Results: KSC-CM increased the proliferation of HFDPCsand ORS cells, and increased the S-phase of the cell cycle in HFDPCs. After subcutaneous injection of KSC-CM in C3H/HeN mice, a significant increase in hair growth and increased proliferation of hair matrix keratinocytes ex vivo was observed. We identified six proteins enriched in KSC-CM. A growth-factor cocktail that contains these six recombinant growth factors significantly increased the proliferation of HFDPCs and ORS cells and enhanced the hair growth of mouse models. Conclusion: These results collectively indicate that KSC-CM has the potential to increase hair growth via the proliferative capacity of HFDPCs and ORS cells.

Growth Inhibition After Exposure to Transforming Growth Factor-β1 in Human Bladder Cancer Cell Lines

이창호,이상한,김두상,전윤수,이남규,이상은 대한비뇨의학회 2014 Investigative and Clinical Urology Vol.55 No.7

Purpose: Transforming growth factor-β1 (TGF-β1) plays a dual role in apoptosis andin proapoptotic responses in the support of survival in a variety of cells. The aim of thisstudy was to determine the function of TGF-β1 in bladder cancer cells. Materials and Methods: The role of TGF-β1 in bladder cancer cells was examined byobserving cell viability by using the tetrazolium dye (MTT) assay after treating the bladdercancer cell lines 253J, 5637, T24, J82, HT1197, and HT1376 with TGF-β1. Amongthese cell lines, the 253J and T24 cell lines were coincubated with TGF-β1 and the pananti-TGF-β antibody. Fluorescence-activated cell sorter (FACS) analysis was performedto determine the mechanism involved after TGF-β1 treatment in 253J cells. Results: All six cell lines showed inhibited cellular growth after TGF-β1 treatment. Although the T24 and J82 cell lines also showed inhibited cellular growth, the growthinhibition was less than that observed in the other 4 cell lines. The addition of pan anti-TGF-β antibodies to the culture media restored the growth properties that had beeninhibited by TGF-β1. FACS analysis was performed in the 253J cells and the 253J cellswith TGF-β1. There were no significant differences in the cell cycle between the twotreatments. However, there were more apoptotic cells in the TGF-β1-treated 253J cells. Conclusions: TGF-β1 did not stimulate cellular proliferation but was a growth inhibitoryfactor in bladder cancer cells. However, the pattern of its effects depended onthe cell line. TGF-β1 achieved growth inhibition by enhancing the level of apoptosis.

Hair-Growth-Promoting Effect of Conditioned Medium of High Integrin α ₆ and Low CD 71 (α ₆ ^bri /CD71 ^dim ) Positive Keratinocyte Cells

Won, Chong Hyun,Jeong, Yun-Mi,Kang, Sangjin,Koo, Tae-Sung,Park, So-Hyun,Park, Ki-Young,Sung, Young-Kwan,Sung, Jong-Hyuk MDPI 2015 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.16 No.3

<P>Keratinocyte stem/progenitor cells (KSCs) reside in the bulge region of the hair follicles and may be involved in hair growth. Hair follicle dermal papilla cells (HFDPCs) and outer root sheath (ORS) cells were treated with conditioned medium (CM) of KSCs. Moreover, the effects of KSC-CM on hair growth were examined <I>ex vivo</I> and <I>in vivo</I>. A human growth factor chip array and RT-PCR were employed to identify enriched proteins in KSC-CM as compared with CM from keratinocytes. KSC-CM significantly increased the proliferation of HFDPCs and ORS cells, and increased the S-phase of the cell cycle in HFDPCs. KSC-CM led to the phosphorylation of ATK and ERK1/2 in both cell types. After subcutaneous injection of KSC-CM in C3H/HeN mice, a significant increase in hair growth and increased proliferation of hair matrix keratinocytes <I>ex vivo</I> was observed. We identified six proteins enriched in KSC-CM (amphiregulin, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-5, granulocyte macrophage-colony stimulating factor, Platelet-derived growth factor-AA, and vascular endothelial growth factor). A growth-factor cocktail that contains these six recombinant growth factors significantly increased the proliferation of HFDPCs and ORS cells and enhanced the hair growth of mouse models. These results collectively indicate that KSC-CM has the potential to increase hair growth via the proliferative capacity of HFDPCs and ORS cells.</P>

조혈세포에 대한 조혈성장인자의 단기간 처리가 CXCR4 발현 및 Stromal Cell-Derived Factor-1 (SDF-1) 매개 경내피세포 이동에 미치는 영향

조덕연,황진희,곽승근,신현영,김성은,윤환중,성주명,김삼용 大韓血液學會 2001 大韓血液學會誌 Vol.36 No.4

Effect to Testosterone on the Growth of Primary Rabbit Proximal Tubule Cells in Serum-Free Medium

추민호,박승준,정주호,정지창,Chu Min-Ho,Park Seung-Joon,Chang Joo-Ho,Jung Jee-Chang The Korean Society of Pharmacology 1995 대한약리학잡지 Vol.31 No.1

Testosterone이 serum-free medium에서 배양한 토끼의 신장 근위세뇨관 상피세포의 세포성장과 기능에 미치는 영향을 관찰한 바 다음과 같은 결과를 얻었다. 1. 토끼의 신장 근위세뇨관 상피세포는 testosterone 1 nM의 농도에서 유의한 세포 성장 촉진 효과를 나타내었고, testosterone 10 nM이상의 농도에서는 세포성장이 억제되었다. 2. Testosterone은 serum-free medium에서 성장촉진인자의 하나인 hydrocortisone을 growth supplement로 넣어준 serum-free medium에서 토끼 신장의 근위세뇨관 상피세포의 성장을 촉진시키었다. 3. Testosterone은 hydrocortisone을 growth supplement로 넣어준 serum-free medium에서 토끼 신장의 근위세뇨관 상피세포의 성장을 촉진시키었다. 4. Testosterone은 Northern blot analysis에 의하여 확인한 토끼 신장의 근위 세뇨관 상피세포의 ${\beta}-actin$ mRNA level은 증가되었다. 이상의 결과로 미루어 보아, serum-free 그리고 hormonally defined media에서 testosterone이 토끼의 신장 근위세뇨관 상피세포의 성장 및 기능에 대하여 촉진적으로 작용하는 것은 cellular mecrofilament의 중요한 구성단백의 하나로 밝혀진 ${\beta}-actin$의 합성 증가에 기인하는 것으로 생각된다. In order to examine the effect of testosterone of the cell growth, using a primary rabbit kidney proximal tubule cell culture system, we observed the effect of 3 growth factors and testosterone supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of testosterone showed a potentiation of the effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (>10 nM) of testosterone indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, testosterone caused to potentiate the growth of the cell. In the presence of hydrocortisone, testosterone also potentiated the grwoth of the proximal tubule cells. According to the Northern analysis, testosterone increased significantly the level of ${\beta}-actin$ mRNA in proximal tubular cells of rabbit kidney. Consequently we may suggest that growth stimulatory effect of testosterone on the primary rabbit kidney proximal tubule cell in serum-free and hormonally defined media ascribed to increase the synthesis of ${\beta}-actin$, which is an important protein consisting of cellular microfilament.