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5. Microcal~lers were allowed to settle for 1 111111.
6. Microcarriers \\ere pellcttcd by sp~iili~iig hr 2 sccuiids ill ii IIIIC~UI'U~I' 111be.
7. Liquid was relllo\ed and dihc;lrded.
S I-IU 111. 01 70'; ctI~;r~~ol \ins ;iddcd \\II~ULII dl\t~irht~lg tll~ pellcl
9. Llquid \\;ij renloicil ;III~ di\cilrticd
lU. l4U 1iL of IOU";
etIl,lilol \\;is ,idilcd \r~tliuut d15turb111g 11111 ~pcllci
1 I, L~ilu~d \+;15 re111oi cci ;111d tl~s~;~rclc~l.
12, 48 } ll~ 01 IOU0" c1l1.111c1l \\$IS ~lllll~~ll.
13. Tile pcllcl \v.i, gellily r11~~1~j)cticIcd b) i;ij)/)i~~g tlie .\liIe 11I'tlic iubc \c\,cr;11 li~llcs, illid tllell
by vorlcxlilg ;it low apecd lbr!-3
\CCOIIL~~.
Si\ allquuta 01.6 pi. c,lcli ul' ~n~croc;irricrs \\ere collcclcd a~rd triln\li.rrcd
to tile center ol'a
~ii;icroc:~rrlcr. Equ,il ;III~O~II~I~(500 big) ol' ~nicriic;irr~crs c;icli IIIII~ ;iiiiI 10 rpl.cad cicllly over
tlic cc1itr~11 I el11 ctr~ 11li11c :I\ ;I g10i11) \\ IIIIIII ;I di;i~i~c~cr 01' I ,5 111~11 Thc la11iiii;ir
hood consislillg ol'll~e biollsl~cs gene g(111 ibil\ 1l1i110~1gIlly CIC~IIIC~I \~ilIr IOUUh ethnnol. The
petri plate colltaii~ing tile cxpla111\ w;is placed ;II ;In ciplxupri;lte d~at:~nce li.oln tl~e device
consistllig of rbe macrocarricr,
The explat)ls iwrc bo~ilbardcd with the lnicrocarriers by
applyllig appropriate pressure ill tlie r,ingc of000 lo I 100 psi. Sprcadilig oftlie iniicrocarriers
was ensured and tlic eupl;l~its \\ere sepa~ited atid \\ere culrurcd 011 shoot induction nlediuin
(SIM) containing h.1S with 4 pM TIIZ, 10 pM 2-IP and 2 pM kinctili. They were incub;lted