View source for 3T3-L1

{{Short description|Cell line used in biological research}}
[[File:Differentiated 3T3-L1 Cell line stained with Oil O Red.jpg|thumb|Differentiated adipocytes in a 3T3-L1 cell line stained with [[Oil Red O]]]]
'''3T3-L1''' is a sub clonal [[cell line]] derived from the original 3T3 Swiss albino cell line of 1962. The 3T3 original cell line was isolated from a mouse embryo and propagated for this specific line of [[3T3 cells]] is used to study [[adipose tissue]]-related diseases and dysfunctions. The 3T3-L1 Swiss subclone line has been widely utilized, since its development, due to its affinity for lipid droplet deposition ''in vitro''. 3T3-L1 cells have a [[fibroblast]]-like [[Morphology (biology)|morphology]], but, under appropriate conditions, the cells [[Cellular differentiation|differentiate]] into an [[adipocyte]]-like [[phenotype]], providing an exemplar model for white adipocytes.<ref>{{cite web |title=3T3-L1 Cell Line |url=http://www.nih3t3.com/transfection-information/ |access-date=22 July 2018}}</ref><ref name=":02">{{Cite journal |last1=Green |first1=Howard |last2=Kehinde |first2=Olaniyi |date=March 1974 |title=Sublines of mouse 3T3 cells that accumulate lipid |url=http://dx.doi.org/10.1016/0092-8674(74)90126-3 |journal=Cell |volume=1 |issue=3 |pages=113–116 |doi=10.1016/0092-8674(74)90126-3 |issn=0092-8674}}</ref><ref name=":12">{{Cite journal |last1=Zebisch |first1=Katja |last2=Voigt |first2=Valerie |last3=Wabitsch |first3=Martin |last4=Brandsch |first4=Matthias |date=June 2012 |title=Protocol for effective differentiation of 3T3-L1 cells to adipocytes |url=http://dx.doi.org/10.1016/j.ab.2012.03.005 |journal=Analytical Biochemistry |volume=425 |issue=1 |pages=88–90 |doi=10.1016/j.ab.2012.03.005 |pmid=22425542 |issn=0003-2697}}</ref> 3T3-L1 cells can be utilized to study a number of cellular and molecular mechanisms related to [[Insulin resistance|insulin-resistance]], [[obesity]], and [[diabetes]] ''in vitro.''<ref name=":02" /> Aside from its usages, this cell line is widely developed and can be purchased for continuous propagation for numerous research studies.<ref>{{Cite journal |last1=Poulos |first1=Sylvia P |last2=Dodson |first2=Michael V |last3=Hausman |first3=Gary J |date=October 2010 |title=Cell line models for differentiation: preadipocytes and adipocytes |url=http://dx.doi.org/10.1258/ebm.2010.010063 |journal=Experimental Biology and Medicine |volume=235 |issue=10 |pages=1185–1193 |doi=10.1258/ebm.2010.010063 |pmid=20864461 |s2cid=24179092 |issn=1535-3702}}</ref> 3T3-L1 cells of the adipocyte morphology increase the synthesis and accumulation of [[triglyceride]]s and acquire the signet ring appearance of adipose cells. These cells are also sensitive to lipogenic and lipolytic hormones, as well as drugs, including [[epinephrine]], [[isoproterenol]], and [[insulin]].<ref name=":42">{{cite journal |author=Green H, Kehinde O |year=1975 |title=An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion |journal=Cell |volume=5 |issue=1 |pages=19–27 |doi=10.1016/0092-8674(75)90087-2 |pmid=165899|s2cid=19040294 }}</ref>
== Lineage development ==
The 3T3-L1 cell line is a sub clone that was initially developed from a mouse embryo, from a clonal expansion of Swiss 3T3 cells.<ref name=":02" /><ref name=":12" /> In 1962, the original [[3T3 cells|3T3 cell]] line that it was established by George Todaro and Howard Green of the [[New York University Grossman School of Medicine|New York University School of Medicine]].<ref name=":52">{{Cite journal |last1=Todaro |first1=George J. |last2=Green |first2=Howard |date=1963-05-01 |title=Quantitative Studies of the Growth of Mouse Embryo Cells in Culture and Their Development Into Established Lines |url=http://dx.doi.org/10.1083/jcb.17.2.299 |journal=Journal of Cell Biology |volume=17 |issue=2 |pages=299–313 |doi=10.1083/jcb.17.2.299 |pmid=13985244 |pmc=2106200 |issn=1540-8140}}</ref> The original cell line was developed through the 3T3 process, which is where the cells derive their naming nomenclature. The 3T3 process began with cultures having 3 days to propagate on a plate (the first "3"), and then a transfer (the "T") of 300,000 cells (second "3") to a new plate to restart the process. These cells will be identified as 3T3 only after they are taken through 20 to 30 passages and have established at a stable growth rate.<ref>{{Cite book |last=Capes-Davis |first=Amanda |url=http://worldcat.org/oclc/1163959689 |title=Freshney's culture of animal cells : a manual of basic technique and specialized applications |date=June 2021 |publisher=Wiley |isbn=978-1-119-51301-8 |oclc=1163959689}}</ref> These cells were originally used to study the properties of transformed cells and the mechanisms of [[neoplastic transformation]]. In 1971, Green and Kehinde established a subline of [[3T3 cells]], called 3T3-L1, which demonstrated a greater propensity for [[Cellular differentiation|differentiation]] into [[adipocyte]]s and a higher affinity for [[lipid]] deposition. The cells were cultured in a [[Fetal bovine serum|serum]]-containing medium and demonstrated a [[fibroblast]]-like morphology. In the early 1980s, it was discovered that these cells could be induced to differentiate into adipocytes in response to hormonal stimulation.<ref name=":02" /><ref name=":42" /><ref name=":62">{{Cite journal |last1=Lane |first1=M. Daniel |last2=Quang |first2=Qi-Qun |date=2005 |title=From multipoint stem cell to adipocyte. |journal=Birth Defects Research Part A: Clinical and Molecular Teratology |volume=73 |issue=7 |pages=476–477|doi=10.1002/bdra.20150 |pmid=15959887 |doi-access=free }}</ref>
== Cell culture characteristics ==
3T3-L1 cells, similar to their other 3T3 counterparts, are typically propagated as an adherent monolayer within a culture vessel. Basal cell culture medias for 3T3-L1 cells tends to contain a version of Dubelcco's Modified Eagle's Medium ([[DMEM]]), [[fetal bovine serum]] (FBS), and an [[antibiotic]] at specific levels dependent upon protocol.<ref>{{Cite journal |last1=Sun |first1=Tingwan |last2=Fu |first2=Mingui |last3=Bookout |first3=Angie L. |last4=Kliewer |first4=Steven A. |last5=Mangelsdorf |first5=David J. |date=2009-06-01 |title=MicroRNA let-7 Regulates 3T3-L1 Adipogenesis |url=http://dx.doi.org/10.1210/me.2008-0298 |journal=Molecular Endocrinology |volume=23 |issue=6 |pages=925–931 |doi=10.1210/me.2008-0298 |pmid=19324969 |pmc=2691679 |issn=0888-8809}}</ref> The DMEM provides essential nutrients, while the FBS provides vital [[growth factor]]s, and the antibiotic protects against culture contamination. The process of [[adipogenesis]], and the start of differentiation can be induced in 3T3-L1 cells through the addition of compounds such as a [[Glucocorticoid|synthetic glucocorticoid]], a [[phosphodiesterase inhibitor]], and [[insulin]]. The most commonly used synthetic [[glucocorticoid]] in adipocyte differentiation procedures tends to be dexamethasone and the most commonly used [[phosphodiesterase inhibitor]] is 1-methyl-3-isobutyl-xanthine ([[IBMX]]). These compounds are usually used in varying combinations and concentrations in differentiation media dependent upon the protocol utilized.<ref name=":12"/><ref>{{Cite journal |last1=Russell |first1=T R |last2=Ho |first2=R |date=December 1976 |title=Conversion of 3T3 fibroblasts into adipose cells: triggering of differentiation by prostaglandin F2alpha and 1-methyl-3-isobutyl xanthine. |journal=Proceedings of the National Academy of Sciences |volume=73 |issue=12 |pages=4516–4520 |doi=10.1073/pnas.73.12.4516 |pmid=188043 |pmc=431523 |bibcode=1976PNAS...73.4516R |issn=0027-8424 |doi-access=free }}</ref><ref>{{Cite journal |last1=Rubin |first1=C.S. |last2=Hirsch |first2=A. |last3=Fung |first3=C. |last4=Rosen |first4=O.M. |date=October 1978 |title=Development of hormone receptors and hormonal responsiveness in vitro. Insulin receptors and insulin sensitivity in the preadipocyte and adipocyte forms of 3T3-L1 cells. |journal=Journal of Biological Chemistry |volume=253 |issue=20 |pages=7570–7578 |doi=10.1016/s0021-9258(17)34541-6 |pmid=81205 |issn=0021-9258|doi-access=free }}</ref><ref name=":22">{{Cite journal |last1=Takenouchi |first1=Takato |last2=Takayama |first2=Yoshiharu |last3=Takezawa |first3=Toshiaki |date=March 2004 |title=Co-treatment with dexamethasone and octanoate induces adipogenesis in 3T3-L1 cells |url=http://dx.doi.org/10.1016/j.cellbi.2003.11.020 |journal=Cell Biology International |volume=28 |issue=3 |pages=209–216 |doi=10.1016/j.cellbi.2003.11.020 |pmid=14984747 |s2cid=25922187 |issn=1065-6995}}</ref><ref name=":32">{{Cite journal |last1=Morrison |first1=Shona |last2=McGee |first2=Sean L |date=2015-04-18 |title=3T3-L1 adipocytes display phenotypic characteristics of multiple adipocyte lineages |url=http://dx.doi.org/10.1080/21623945.2015.1040612 |journal=Adipocyte |volume=4 |issue=4 |pages=295–302 |doi=10.1080/21623945.2015.1040612 |pmid=26451286 |pmc=4573194 |issn=2162-3945}}</ref> 3T3-L1 cells are commonly utilized as an effective model for [[white adipocyte]]s because of their similar [[fibroblast]]-like [[Morphology (biology)|morphology]], lipid accumulation, [[gene expression]], and basal energy exchanges and transformations.<ref name=":32" /> Although, some literature suggests that 3T3-L1 adipocytes can possess certain [[Phenotype|phenotypic]] signatures similar to other adipocyte lineages aside from white adipocytes.<ref name=":32" />
== Adipogenesis and lipogenesis ==
[[Adipogenesis]] can be described as the effective differentiation of preadipocytes into mature adipocytes that can then undergo lipogenesis. During this period, cells can undergo hyperplastic growth until they are differentiated. Since 3T3-L1 cells are an [[Immortalised cell line|immortalized cell line]] and require a hormonal differentiation, there has been much debate on their comparability to ''in vivo'' studies and [[Primary cell culture|primary cell lines]]. Aside from the synthetic differentiation itself, 3T3-L1 lineages can display low differentiation efficiency when utilizing common differentiation methods. Low differentiation efficiency can change [[adipogenesis]] and [[lipogenesis]] related experiment results and limit result interpretation. Studies suggest that differentiation efficiency can rely on factors such as [[culture dish]] material, culture dish provider, culture dish type, cell confluence at the time of differentiation.<ref name=":12" /><ref>{{Cite journal |last1=Mehra |first1=Anisha |last2=Macdonald |first2=Ian |last3=Pillay |first3=Tahir S. |date=March 2007 |title=Variability in 3T3-L1 adipocyte differentiation depending on cell culture dish |url=http://dx.doi.org/10.1016/j.ab.2006.12.016 |journal=Analytical Biochemistry |volume=362 |issue=2 |pages=281–283 |doi=10.1016/j.ab.2006.12.016 |pmid=17241610 |issn=0003-2697}}</ref> [[Lipogenesis]] can be described as the biochemical and physical accumulation of [[triacylglyceride]]s in a differentiated adipocyte. Again, the deposition of [[adipose tissue]] relies on the number of [[adipocyte]]s, their differentiation efficiency and overall lipid accumulation. It is not unrelated that the inhibition of one of these processes could impact others as well.
== Advantages and disadvantages ==
One advantage of using 3T3-L1 cells is their well-characterized [[Cellular differentiation|differentiation]] process, which allows for reproducible experiments and comparison of results across studies.<ref name=":52" /><ref name=":62" /> Additionally, the cells can be easily cultured and maintained in the laboratory, and are relatively inexpensive compared to other [[Immortalised cell line|cell lines]] or [[Model organism|animal models]]. However, one limitation of using 3T3-L1 cells is that they are derived from mice and may not fully recapitulate [[human biology]]. Additionally, 3T3-L1 cells are a [[Homogeneity and heterogeneity|homogeneous]] cell population, which may not fully reflect the [[Homogeneity and heterogeneity|heterogeneity]] of [[adipose tissue]] [[in vivo]].<ref name=":62" /> These limitations can affect comparability of this particular cell lineage.
== Discoveries ==
While 3T3-L1 gene expression mimics that of a white adipocyte, some literature suggests some phenotypic 3T3-L1 characteristics can resemble that of brown adipocytes. 3T3-L1 cells, when supplemented with catecholamines, utilized [[Thermogenin|UCP-1]] to increase oxygen consumption similar to [[Brown adipose tissue|brown adipocytes.]] This brown adipocyte characteristic was only enhanced with long-term [[catecholamine]] supplementation.<ref name=":32" /> While the 3T3-L1 lineage can display characteristics similar to both white and [[Brown adipose tissue|brown adipocytes]], they have had different results in relation to beige adipocyte [[phenotype]]<nowiki/>characteristics. When 3T3-L1 cells are differentiated into beige adipocytes, they fail to express any beige phenotypic signatures.<ref name=":32" /> The process of differentiation is constantly being re-evaluated in the adipose research sector. While a majority of ''in vitro'' experiments utilize the common [[insulin]], dexamethasone, and [[IBMX]] combinations, others have found other methods of differentiating and inducing the process of adipogenesis. Other methods can include transfection with a [[Ras GTPase|''ras'' oncogene]], the combination of dexamethasone and [[Caprylic acid|octanoate]], and the normal combination with the addition of a [[PPAR agonist|peroxisome proliferator-activated receptor y (PPARy) agonist]].<ref name=":12" /><ref name=":22" /><ref>{{Cite journal |last1=Benito |first1=Manuel |last2=Porras |first2=Almudena |last3=Nebreda |first3=Angel R. |last4=Santos |first4=Eugenio |date=1991-08-02 |title=Differentiation of 3T3-L1 Fibroblasts to Adipocytes Induced by Transfection of ''ras'' Oncogenes |url=http://dx.doi.org/10.1126/science.1857988 |journal=Science |volume=253 |issue=5019 |pages=565–568 |doi=10.1126/science.1857988 |pmid=1857988 |bibcode=1991Sci...253..565B |issn=0036-8075}}</ref> While some protocols promote the process of adipogenesis in 3T3-L1 cells, others reduce or inhibit the process. In 3T3-L1 adipocytes, oleanolic acid (5''u''mol/L) down regulated the expression of [[Peroxisome proliferator-activated receptor gamma|PPARy]] and [[CCAAT/enhancer-binding protein alpha|cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT) enhancer binding protein ''a'']] (C/EBP''a'') during differentiation. Thus, [[oleanolic acid]] suppressed adipocyte differentiation and consequential [[lipid]] accumulation when applied in differentiation media to 3T3-L1 cells. [[Berberine]] impacted [[GATA2|GATA-2]] and [[GATA3|GATA-3]] and thus inhibited 3T3-L1 differentiation when applied during differentiation or post-differentiation ''in vitro''.<ref>{{Cite journal |last1=Hu |first1=Y. |last2=Davies |first2=G.E. |date=September 2009 |title=Berberine increases expression of GATA-2 and GATA-3 during inhibition of adipocyte differentiation |url=http://dx.doi.org/10.1016/j.phymed.2009.03.002 |journal=Phytomedicine |volume=16 |issue=9 |pages=864–873 |doi=10.1016/j.phymed.2009.03.002 |pmid=19403287 |issn=0944-7113}}</ref> Long term treatment of [[flavonoid]], [[luteolin]], affected PPARy activation and suppressed [[adipogenesis]] 3T3-L1 adipocytes.<ref>{{Cite journal |last1=Park |first1=Hee-Sook |last2=Kim |first2=Soon-Hee |last3=Kim |first3=Young Sup |last4=Ryu |first4=Shi Yong |last5=Hwang |first5=Jin-Taek |last6=Yang |first6=Hye Jeong |last7=Kim |first7=Gun-Hee |last8=Kwon |first8=Dae Young |last9=Kim |first9=Myung-Sunny |date=July 2009 |title=Luteolin inhibits adipogenic differentiation by regulating PPARγ activation |url=http://dx.doi.org/10.1002/biof.38 |journal=BioFactors |volume=35 |issue=4 |pages=373–379 |doi=10.1002/biof.38 |pmid=19353690 |s2cid=30777034 |issn=0951-6433}}</ref> Isopanduratin A also inhibited adipogenesis in 3T3-L1 adipocytes by impacting multiple targets in the adipogenic growth cycle such as [[Peroxisome proliferator-activated receptor gamma|PPARy]], [[CEBPA|C/EBP''a'']], sterol regulatory binding protein- 1c ([[Sterol regulatory element-binding protein 1|SREB-1c]]), [[adiponectin]], and [[lipoprotein lipase]](LPL).<ref>{{Cite journal |last1=Rungsa |first1=Prapenpuksiri |last2=San |first2=Htoo Tint |last3=Sritularak |first3=Boonchoo |last4=Böttcher |first4=Chotima |last5=Prompetchara |first5=Eakachai |last6=Chaotham |first6=Chatchai |last7=Likhitwitayawuid |first7=Kittisak |date=2023-02-27 |title=Inhibitory Effect of Isopanduratin A on Adipogenesis: A Study of Possible Mechanisms |journal=Foods |language=en |volume=12 |issue=5 |pages=1014 |doi=10.3390/foods12051014 |issn=2304-8158 |pmc=10000982 |pmid=36900533 |doi-access=free }}</ref>
==Sources==
{{reflist}}
==External links==
*[https://www.cellosaurus.org/CVCL_0123 Cellosaurus entry for 3T3-L1]
[[Category:Rodent cell lines]]
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