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s A c v e w s c f s o g c c o A c M o c s f w T s t a a L. tarentolae suspension c performed a tarentolae w All growth m Leishmania. completion w vitality the c exceeded, it weeks For mainta suspension c culture (TC) f faster. It is no Best results successive d occasional h growth. It is incubate TC cultivate Leis cultivation fo of the LEXSY Always contr cultures are o motile. Thes Mid-growth p of different s containing s shape) and limitations a fungal or oth Keep patient with regular They need th If you - desp sediment cel incubation in transfection Don’t centrifu by rigorous v at 2000-300 pellets easier If you cultiva aerate the cu 10 9 cells/ml. How e needs ae culture with re t 26°C. Hig ill not survive media should Hemin is l with Hemin th completed m t is possible aining LEXSY cultures in 10 flasks. Don´t ot necessary are obtaine dilution of c higher dilutio convenient t flask uprig shmania muc or transfection manuals rol appearan of drop-like s se cells are phase culture shape. Don’t such subpop remain mot nd must not her contamina t, esp. if you doubling tim heir time to g pite following ls 3 min at 2 n ventilated T uge Leishma vortexing. Th 00g is sufficie r. If required, ate LEXSY str ulture in a 1 . If you intend Jena Bioscien w to grow erobic cond egular dilutio her temperat e at 37°C be suppleme ight-sensitive he medium m edium shoul to re-add H Y strains for 0 ml LEXSY use agitated or growth-pr ed with inoc cultures of lo ns of stationa to dilute 10 ht, lowering ch longer tha n see chapte nce and mot shape, appro e most effic es always con t hesitate to pulations. Ce tile. Enhance necessarily ations may b u are used to mes of 7 h in row or to ad g these instr 2000g, resus TC flasks. Thi nia cultures he cells are s ent for sedim , prolong cen rains in bior 0 L fermenta d to use a sti nce GmbH • Loebstedte Tel.: +4 w a Leish ditions. The ons (see chap tures lower th ented with H , so Leishm must be store d be used w Hemin (and P transfection BHI mediu d cultures for romoting, to a culations dur ow cell den ary cells at e ml cultures g aeration fo an for 7 da r 4.4, for cul ility of the ce ox. 15x5 μm cient for tra ntain subpop transfect, p ells of older ed motility m be a sign o be identified b o working w n static suspe apt to new c uctions - enc spend pellet c is approach at high spee sensitive to th mentation and ntrifugation ti reactors be c ation without rrer, avoid h er Str. 80 • D-07749 J 49-3641-6285000 • F hmania cu strains can pter 3.2 of LE he growth-ra Hemin (Cat. mania must b ed in the dar within 2 wee PenStrep) and and analys m (Cat. No. r strain main add serum to ring early st nsities as th e.g. 1:100 w 1:50 on Mo or longer in ays in the sa ltivation for p ells by micro in size with ansfection pulations of n late or prese cultures get may result fr of mid grow by microscop with bacteria. ension culture onditions counter grow carefully in f was very hel ed >10.000g hese procedu d makes gen ime rather tha careful with stirring for o high sharing f Jena • Germany • e-M Fax: +49-3641-62851 ulture n be mainta EXSY manual tes and vital No ML-108) be cultivated k at 4°C. Fo eks. Howeve d to use this sis it is conv . ML-411, 41 tenance sinc o the BHI med tationary ph is may dro will not advers nday and 1 tervals betw ame medium protein expre oscopy. Cells one flagellu and platin non- or less m erve a cultur t longer and rom nutrient wth culture st py . Leishmania es and 4-5 h wth problems resh growth lpful in rescu g and don’t ures and ma ntle and quic an using hig stirring. We obtaining hig forces Mail: info@jenabioscien 0 ained as co ls). All cultiva ity significan which is ess d in the da or optimal gro er, if this she s medium fo venient to gr 12) in ventila ce cells will a dium ase. Avoid p growth. H sely affect su :20 on Frida ween passage without dilu ession see cha of mid-grow m at the flat ng on solid motile cells an re with drop d thinner (n deprivation tage. Also, a cells are pr h in agitated s with the ho medium and uing cultures resuspend c ay lyse. Cent k resuspensio her speed e found it su gh cell densi nce.com ontinuous ations are tly and L. sential for ark. After owth and elf live is r 2 more row static ted tissue age much repeated However, ubsequent ay and to es. Don’t ution. For apter 5.4 wth phase end, and d media. nd of cells p-like cells eedle-like or other bacterial, rotozoans d cultures. ost strain, d continue esp. after ell pellets rifugation on of cell fficient to ties up to

How to grow a Leishmania culture - Jena Bioscience is l ith Hemin th ompleted m is possible ining LEXSY ultures in 10 lasks. Don´t t necessary are obtaine ilution of c igher dilutio

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Page 1: How to grow a Leishmania culture - Jena Bioscience is l ith Hemin th ompleted m is possible ining LEXSY ultures in 10 lasks. Don´t t necessary are obtaine ilution of c igher dilutio

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L. tarentolaesuspension cperformed atarentolae w

All growth mLeishmania. completion wvitality the cexceeded, itweeks

For maintasuspension cculture (TC) ffaster. It is no

Best results successive doccasional hgrowth. It is incubate TCcultivate Leiscultivation foof the LEXSY

Always contrcultures are omotile. ThesMid-growth pof different scontaining sshape) and limitations afungal or oth

Keep patientwith regular They need th

If you - despsediment celincubation intransfection

Don’t centrifuby rigorous vat 2000-300pellets easier

If you cultivaaerate the cu109 cells/ml.

How

e needs aeculture with ret 26°C. Higill not survive

media should Hemin is l

with Hemin thcompleted mt is possible

aining LEXSYcultures in 10flasks. Don´t ot necessary

are obtainedilution of chigher dilutio

convenient t flask uprigshmania mucor transfection

manuals

rol appearanof drop-like sse cells arephase cultureshape. Don’tsuch subpopremain motnd must not

her contamina

t, esp. if youdoubling tim

heir time to g

pite followingls 3 min at 2n ventilated T

uge Leishmavortexing. Th00g is sufficier. If required,

ate LEXSY strulture in a 1. If you intend

Jena Bioscien

w to grow

erobic condegular dilutioher temperat

e at 37°C

be supplemeight-sensitivehe medium medium shoulto re-add H

Y strains for 0 ml LEXSY

use agitatedor growth-pr

ed with inoccultures of lons of stationato dilute 10 ht, loweringch longer than see chapte

nce and motshape, approe most effices always cont hesitate to

pulations. Cetile. Enhance

necessarily ations may b

u are used tomes of 7 h inrow or to ad

g these instr2000g, resusTC flasks. Thi

nia cultures he cells are sent for sedim, prolong cen

rains in bior0 L fermentad to use a sti

nce GmbH • LoebstedteTel.: +4

w a Leish

ditions. The ons (see chaptures lower th

ented with H, so Leishm

must be stored be used w

Hemin (and P

transfectionBHI mediud cultures forromoting, to a

culations durow cell denary cells at eml cultures

g aeration foan for 7 dar 4.4, for cul

ility of the ceox. 15x5 μm cient for trantain subpoptransfect, p

ells of older ed motility m

be a sign obe identified b

o working wn static suspeapt to new c

uctions - encspend pellet cis approach

at high speesensitive to thmentation andntrifugation ti

reactors be cation without rrer, avoid h

er Str. 80 • D-07749 J49-3641-6285000 • F

hmania cu

strains canpter 3.2 of LEhe growth-ra

Hemin (Cat. mania must bed in the darwithin 2 weePenStrep) and

and analysm (Cat. No.r strain mainadd serum to

ring early stnsities as the.g. 1:100 w1:50 on Moor longer inays in the saltivation for p

ells by microin size with

ansfection pulations of nlate or presecultures get

may result frof mid growby microscop

with bacteria.ension cultureonditions

counter growcarefully in fwas very hel

ed >10.000ghese procedud makes genime rather tha

careful with stirring for o

high sharing f

Jena • Germany • e-MFax: +49-3641-62851

ulture

n be maintaEXSY manualtes and vital

No ML-108)be cultivatedk at 4°C. Foeks. Howeved to use this

sis it is conv. ML-411, 41tenance sinc

o the BHI med

tationary phis may dro

will not adversnday and 1tervals betw

ame medium protein expre

oscopy. Cellsone flagelluand platin

non- or less merve a culturt longer androm nutrient wth culture stpy

. Leishmaniaes and 4-5 h

wth problemsresh growth lpful in rescu

g and don’t ures and mantle and quican using hig

stirring. Weobtaining higforces

Mail: info@jenabioscien0

ained as cols). All cultivaity significan

which is essd in the daor optimal groer, if this shes medium fo

venient to gr12) in ventilace cells will adium

ase. Avoid p growth. Hsely affect su:20 on Frida

ween passagewithout dilu

ession see cha

of mid-growm at the flat

ng on solidmotile cells anre with dropd thinner (ndeprivation

tage. Also,

a cells are prh in agitated

s with the homedium and

uing cultures

resuspend cay lyse. Centk resuspensioher speed

e found it sugh cell densi

nce.com

ontinuous ations are tly and L.

sential for ark. After owth and elf live is r 2 more

row static ted tissue

age much

repeated However, ubsequent ay and to es. Don’t ution. For apter 5.4

wth phase end, and

d media. nd of cells p-like cells eedle-like or other

bacterial,

rotozoans d cultures.

ost strain, d continue esp. after

ell pellets rifugation on of cell

fficient to ties up to