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n cultura in vitro i cre terea de organe sau p r i ale acestora, n care diversele componente ale esutului lor, de exemplu,

parenchimului si stroma, sunt p strate att n ceea ce prive te structura lor i func iile, astfel nct organele de cultura se aseam n ndeaproape organele n cauz n vivo se numeste cultura de organe

FEATURES CARACTERISTICI
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New growth occurs in form of differentiated structures. Nou cre tere are loc n form de structuri diferen iate. eg- glandular structure is retained in glands. de exemplu, structura-glandular este re inut n glande. Cultured organ retains its physiological features. Organ de cultur i p streaz caracteristicile sale fiziologice. eg- hormone dependent organ continue to be dependent. de exemplu, hormon-dependente de organe continu s fie dependente. Morphogenesis in cultured fetal tissues ia more or less comparable to that in vivo. Morfogeneza n esuturile fetale cultur ia mai mult sau mai pu in comparabil cu cea in vivo. Outgrowth of isolated cells from periphery of explants can be minimized by manipulating the culture. Excrescen de celule izolate de la periferie de explantelor pot fi minimizate prin manipularea cultura. conditions Condi ii

ADVANTAGES AVANTAJE
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Explant remain comparable to the in vivo organs both in structure & function,developmental of fetal organ is comparable, Explant s r mn comparabile cu organe n vivo, att n structura i func ia, de dezvoltare de organe fetale este comparabil , It provides information on the pattern s of growth, differentiation, & development of an organ. Acesta ofer informa ii cu privire la e modelul de cre tere, diferen iere, i de dezvoltare a unui organ. In some cases, it can replace whole animals in experimentation as the results from are easier to interpret. n unele cazuri, se poate nlocui animale ntregi, n experimentare, deoarece rezultatele de la sunt mai u or de interpretat.

LIMITATIONS LIMIT RI
Results from organ cultures are often not comparable to those from whole animals studies, eg in studies on drug action, since the drug are metabolized in vivo but not in vitro Rezultate ob inute din culturi de organe de multe ori nu sunt comparabile cu cele din studii ntregi animale, de exemplu, n studiile privind ac iunea de droguri, deoarece de droguri sunt metabolizate in vivo, dar nu in vitro

TECHNIQUES OF ORGAN CULTURE: TEHNICI DE CULTUR DE ORGANE: Plasma Clot Method:- The explant is culture on the surface of a clot consisting of chick (or other) plasma & chick embryo extract contained in a watch glass. Plasma Metoda Clot: - explant este cultura pe suprafa a unui cheag de compus de pui (sau de alt natur ) cu
plasm i de embrioni de pui extract de con inut ntr-o sticl de ceas. The watch glass may or may not be closed with a glass lid & sealed with paraffin wax. Sticl de ceas poate sau nu

poate fi nchis cu un capac de sticl i sigilat cu cear de parafin . This has been the classical technique for studying the morphogenesis in embryonic organ rudiments. Acest lucru a fost tehnica clasica pentru studierea morfogeneza n rudimente de organe embrionare. It has been also modified to study the action of hormones, vitamins carcinogens etc on the adult mammalian tissues. Acesta a fost, de asemenea, modificat pentru a studia ac iunea de hormoni, vitamine etc agen i cancerigeni asupra tesuturilor adul i mamifere.

Raft Method:- The explant is placed on to a raft of lens paper or rayon acetate, which is floated on a serum in a watch glass. Metoda Raft: - explant este plasat pe un set de lentile
de hrtie sau de acetat de raion, care este plutit pe un ser ntr-o sticl de ceas. Rayon acetate rafts were made to float on the serum by treating their 4 corners by silicone. Raionul de acetat de plute de salvare s-au f cut s pluteasc pe ser prin tratarea lor 4 col uri de silicon. Similarly, the floatability of lens paper is enhanced by treating with silicone. n mod similar, flotabilitatea de hrtie cristalinului este mbun t it prin tratarea cu silicon. on each raft 4 or more explants are placed. pe fiecare plut 4 sau mai multe explantelor sunt plasate. In a combination of raft and clot techniques, the explant are first placed on a suitable raft, which is then kept on a plasma clot. ntr-o combina ie de tehnici de plut i cheag, explant sunt introduse pentru prima dat pe o plut adecvat, care este apoi men inut la un cheag de plasma. This modification makes media changes easy& prevents the sinking into the liquefied plasma. Aceast modificare face schimb ri mass-media u or i previne scufundarea n plasm lichefiat.

Agar Gel Method:- The medium( consisting of suitable salt solution, serum, chick embryo extract or a mixture of certain amino acids & vitamins) is gelled with 1% agar. Gel Metoda Agar: - mediu (constnd din solu ie sare potrivita, ser, extract de pui embrion sau un amestec
de anumiti aminoacizi i vitamine) este gelifiat cu 1% agar-agar. This method avoids the immersion of explants into the medium & permits the use of defined media. Aceast metod evit imersiune a explantelor n mediu i permite utilizarea de mass-media definite. generally the explants need to be subculutured on fresh agar gels every 5-7 days. n general, explantelor trebuie s fie subculutured pe geluri agar-agar proaspete la fiecare 5-7 zile. The agar gels are generally kept in embryological watch glasses & sealed with paraffin wax. Geluri agar-agar, n general, sunt inute n pahare ceas embriologice i sigilat cu cear de parafin . This method is used to study many developmental aspects of normal organs as well as tumors. Aceast metod este folosit pentru a studia multe aspecte de dezvoltare de organe normale, precum i tumori.

Grid Method:- Devised by trowell in 1954, this method utilizes 25 mm X 25 mm pieces of


suitable wiremesh or pwerforated stainless steelsheet whose edges are bent to form4 legs of about 4 mm height. Metoda Grid: - conceput de trowell n 1954, aceast metod folose te 25 mm X 25 mm, buc i de maro adecvate sau steelsheet pwerforated inoxidabil ale c ror margini sunt ndoite la form4 picioare de aproximativ 4 mm n l ime. Skeletal tissues are generally placed directly on the grid but softer tissues like glandsor skin are vfirst placed on rafts, which are then kept on the grids. esuturile osoase sunt, n general plasate direct pe grila, dar tesuturile moi cum ar fi pielea glandsor vfirst sunt introduse pe plute, care sunt apoi p strate pe grile. the grid themselves are then placed on a culturechamber filled with fluid medium upto the grid; the chamber is supplied witth a mixture of O 2 & Co 2 to meet the high O 2 requirement of adult mammalian organs. grila de sine sunt apoi plasate pe o culturechamber umplut cu mediu lichid pana re elei; camera este furnizat witth un amestec de O 2 i CO 2 pentru a satisface cerin a de nalt O 2 de organe adul i mamifere

Cultura de organe este o dezvoltare de la cultura de tesut metode de cercetare, cultura organ este capabil de a modela cu exactitate func iile de membru al unui organ n diverse state i condi iile de utilizare a efectiv n organul vitro sine. Parts of an organ or a whole organ can be cultured in vitro . Piese de schimb al unui organ sau a unui organ ntregi pot fi cultivate in vitro . The main objective is to maintain the architecture of the tissue and direct it towards normal development. Obiectivul principal este de a men ine arhitectura a esutului i s o dirijeze spre o dezvoltare normala. In this technique, it is essential that the tissue is never be disrupted or damaged. In aceasta tehnica, este esen ial ca tesutul nu este niciodat s fie perturbat sau deteriorate. It thus requires careful handling. Este nevoie de astfel de manipulare atent . The media used for a growing organ culture are generally the same as those used for tissue culture. Mass-media utilizate pentru o cultur de organe de cultur sunt, n general, acelea i ca i cele utilizate pentru cultura de tesut. The techniques for organ culture can be classified into (i) those employing a solid medium and (ii) those employing liquid medium. Tehnicile de cultura de organe pot fi clasificate n (i) cele care utilizeaz un mediu solid i (ii) cele care utilizeaz mediu lichid.

Contents Con inut


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1 Methodology 1 Metodologia o 1.1 Plasma clot method 1.1 Plasma cheag de metoda o 1.2 Agar gel method 1.2 Agar gel metoda  1.2.1 Raft Methods 1.2.1 Metode de Raft  1.2.2 Grid Method 1.2.2 Metoda Grid 2 Limitations 2 Limit ri 3 Current progress 3 curent progrese 4 See also 4 A se vedea, de asemenea, 5 References 5 Referin e 6 External links 6 Leg turi externe

[ edit ] Methodology [ edit ] Metodologia


Embryonic organ culture is an easier alternative to normal organ culture derived from adult animals. Cultura de organe embrionare este o alternativ mai u or de cultura de organe normale provenite de la animalele adulte. The following are three techniques employed for embryonic organ culture. Urm toarele trei tehnici sunt utilizate pentru cultura de organe embrionare.

[ edit ] Plasma clot method [ edit ] Plasma metoda cheag


The following are general steps in organ culture on plasma clots. Urm toarele sunt pa ii generale n cultur de organe de la plasma cheaguri.

1. Prepare a plasma clot by mixing 15 drops of plasma with five drops of embryo extract in a watch glass. Se prepar un cheag de plasma prin amestecarea a 15 pic turi de plasm cu cinci pic turi de extract de embrioni ntr-o sticl de ceas. 2. Place a watch glass on a pad of cotton wool in a petri dish; cotton wool is kept moist to prevent excessive evaporation from the dish. Posta i o sticl de ceas pe un tampon de vat ntr-un vas Petri, vat este mentinut umed pentru a preveni evaporarea excesiv din vas. 3. Place a small, carefully dissected piece of tissue on top of the plasma clots in watch glass. Se pune o bucata mica, cu aten ie i disecat de tesut pe partea de sus a cheagurilor de plasm n sticla de ceas. The technique has now been modified, and a raft of lens paper or rayon net is used on which the tissue is placed. Tehnica a fost acum modificat, i o serie de lentile de hrtie sau plas raional este utilizat pe care tesutul este plasat. Transfer of the tissue can then be achieved by raft easily. Transferul de tesut poate fi apoi realizate de plut cu u urin . Excessive fluid is removed and the net with the tissue placed again on the fresh pool of medium.Parts of an organ or a whole organ can be cultured in vitro. Excesiv de lichide este eliminat i net cu esutul plasat din nou pe bazinul cu apa de medium.Parts al unui organ sau a unui organ ntregi pot fi cultivate in vitro. The main objective is to maintain the architecture of the tissue and direct it towards normal development. Obiectivul principal este de a men ine arhitectura a esutului i s o dirijeze spre o dezvoltare normala. In this technique, it is essential that the tissue never be disrupted or damaged. In aceasta tehnica, este esen ial ca niciodat esuturi s fie perturbate sau deteriorat. It thus requires careful handling. Este nevoie de astfel de manipulare atent . The media used for a growing organ are generally the same as those used for tissue culture. Mass-media utilizate pentru un organ de cultur sunt, n general, acelea i ca i cele utilizate pentru cultura de tesut. The techniques for organ culture can be classified into (i) those employing a solid medium and (ii) those employing liquid medium. Tehnicile de cultura de organe pot fi clasificate n (i) cele care utilizeaz un mediu solid i (ii) cele care utilizeaz mediu lichid.

[ edit ] Agar gel method [ edit ] Metoda de gel de agar


Media solidified with agar are also used for organ culture and these media consist of 7 parts 1% agar in BSS, 3 parts chick embryo extract and 3 parts of horse serum. Media solidificat cu agar-agar sunt, de asemenea, utilizate pentru cultura de organe i de aceste mass-media const din 7 piese de agar-agar de 1% n BSS, 3 piese de embrioni extract de pui de g in i 3 p r i din ser de la cal. Defined media with or without serum are also used with agar. Mass-media definite, cu sau f r ser sunt de asemenea folosite cu agar-agar. The medium with agar provides the mechanical support for organ culture. Mediul cu geloz ofer suport mecanic pentru cultura de organe. It does not liquefy. Ea nu se lichefieze. Embryonic organs generally grow well on agar, but adult organ culture will not survive on this medium. Organe embrionare, n general, cresc bine pe agar-agar, dar cultura de organe obscen nu vor supravie ui pe acest mediu. The culture of adult organs or parts from adult animals is more difficult due to their greater requirement of oxygen . Cultura de organe obscen sau a unor p r i de la animale adulte este mult mai dificil din cauza cerin a lor mai mare de oxigen . A variety of adult organs (eg the liver ) have been cultured using special media with special apparatus (Towell's II culture chamber). O varietate de organe pentru adul i (de exemplu, ficat ) au fost cultivate cu ajutorul mass-media cu aparate speciale de construc ii (Towell a II camera de cultur ). Since serum

was found to be toxic, serum-free media were used, and the special apparatus permitted the use of 95% oxygen. ntruct serice sa dovedit a fi toxice, ser-media libere au fost utilizate, precum i aparate speciale permis utilizarea de 95% oxigen. [ edit ] Raft Methods [ edit ] Raft Metode In this approach the explant is placed onto a raft of lens paper or rayon acetate, which is floated on serum in a watch glass. n aceast abordare explant este plasat pe o serie de lentile de hrtie sau de acetat de raion, care este listat pe ser ntr-o sticl de ceas. Rayon acetate rafts are made to float on the serum by treating their 4 corners with silicone. Raionul de acetat de plute de salvare sunt facute pentru a pluti pe ser prin tratarea lor 4 col uri cu silicon. Similarly, floatability of lens paper is enhanced by treating it with silicone. n mod similar, flotabilitatea de hrtie obiectivului este consolidat prin tratarea cu silicon. On each raft, 4 or more explants are usually placed. La fiecare plut , 4 sau mai multe explantelor sunt de obicei plasate. In a combination of raft and clot techniques, the explants are first placed on a suitable raft, which is then kept on a plasma clot. ntr-o combina ie de tehnici de plut i cheag, explantelor sunt introduse pentru prima dat pe o plut adecvat, care este apoi men inut la un cheag de plasma. This modification makes media changes easy, and prevents the sinking of explants into liquefied plasma. Aceast modificare face ca schimbarea u oar a mass-media, i previne scufundarea explantelor lichefiat n plasm . [ edit ] Grid Method [ edit ] Metoda Grid Initially devised by Trowell in 1954, the grid method utilizes 25 mm x 25 mm pieces of a suitable wire mesh or perforated stainless steel sheet whose edges are bent to form 4 legs of about 4 mm height. Ini ial conceput de Trowell n 1954, metoda de re ea folose te 25 mm x 25 mm, buc i de plas de srm adecvat sau tabla perforata otel inoxidabil ale c ror margini sunt ndoite pentru a forma 4 picioare de aproximativ 4 mm n l ime. Skeletal tissues are generally placed directly on the grid but softer tissues like glands or skin are first placed on rafts, which are then kept on the grids. esuturile osoase sunt, n general plasate direct pe grila, dar tesuturile moi cum ar fi glandele pielii sau sunt introduse pentru prima dat pe plute, care sunt apoi p strate pe grile. The grids themselves are placed in a culture chamber filled with fluid medium up to the grid; the chamber is supplied with a mixture of O 2 and CO 2 to meet the high O 2 requirements of adult mammalian organs. Grile n sine sunt plasate ntr-o camer de cultur umplut cu mediu lichid pn la re ea; camera este livrat cu un amestec de O 2 i CO 2 pentru a satisface cerin ele ridicate de O 2 de organe adul i mamifere. A modification of the original grid method is widely used to study the growth and differentiation of adult and embryonic tissues. O modificare a metodei grilei original este utilizat pe scar larg pentru a studia cresterea si diferentierea de esuturi adulte i embrionare.

[ edit ] Limitations [ edit ] Limit ri


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Results from organ cultures are often not comparable to those from whole animals studies, eg in studies on drug action since the drugs are metabolized in vivo but not in

vitro. Rezultate ob inute din culturi de organe de multe ori nu sunt comparabile cu cele din studii ntregi animale, de exemplu, n studiile privind ac iunea de droguri, deoarece medicamentele sunt metabolizate in vivo, dar nu in vitro.

[ edit ] Current progress [ edit ] progresul curent


In April 2006, scientists reported a successful trial of seven bladders grown in-vitro and given to humans. [ 1 ] A bladder has been cultured by Anthony Atala of the Wake Forest Institute for Regenerative Medicine in Winston-Salem, North Carolina. n aprilie 2006, oamenii de stiinta au raportat un proces de succes de apte vezici cultivate in vitro i a dat la oameni. [1] Un vezicii urinare a fost cultivat de c tre Anthony Atala din Wake Forest Institutul de Medicina Regenerativa din Winston-Salem, Carolina de Nord. A jawbone has been cultured at Columbia University, a lung has been cultured at Yale. O falc a fost cultivati de la Universitatea Columbia, un pl mn a fost cultivati de la Yale. A beating rat heart has been cultured by Doris Taylor at the University of Minnesota. O inim bate obolan a fost cultivate de c tre Doris Taylor de la Universitatea din Minnesota. An artificial kidney has been cultured by H. Un rinichi artificial a fost cultivate de c tre H. David Humes at the University of Michigan. [ 2 ] David Humes de la Universitatea din Michigan.

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